Tecan Journal Edition 02/2018

Tecan Journal

Edition 2/2018

Life Sciences, Diagnostics and Partnering

Building blocks for custom solutions

Redesigning the drug safety workflow

Cell counting made easy

Long-term partners in success

Pages 8-9

Pages 10-11

Pages 16-17

Pages 28-29

CEO WELCOME

Welcome Dear Reader, Automation is increasingly being seen as a ‘must have’ tool in the laboratory, not only because of the higher throughput it offers and the staff time it frees up, but also because of the improvements in precision and reproducibility it represents. This is especially true for regulated labs, where the shift in approach means that more and more laboratories with little or no previous experience using automation are choosing these technologies to accelerate and simplify their workflows. Automation is a particular strength of Tecan’s; through Tecan Partnering we supply more than 35 different diagnostic instrument platforms to more than 30 customers under their own labels, from leading diagnostics companies to segment specialists and emerging players. You can read more about companies following this trend in the Synchron (pages 8-9) and Vircell (pages 28-29) articles in this issue.

Tecan Partnering has a broad and growing portfolio of OEM liquid handling options for the IVD industry, now including the exciting new Cavro® Magni Flex*, a modular, scalable, multichannel XYZ technology platform packed with features to make it easier for instrument designers to cost-effectively integrate advanced functions into their systems. We’re also taking a fresh approach to software development, with the introduction of MAPlinx™*, our modular application platform for OEM customers. This development framework combines standardized modules for functionality common to multiple systems with the ability to create custom application software accelerating development and time to market.

We’ve news too of the end-user business where, following the introduction of the Fluent® Gx for regulated markets in the first regions in February 2018, we also registered the platform with the US FDA as a Class 1 Medical Device in April. Since the launch of the Fluent Gx, we have seen great demand for the newest liquid handling platform on the market, with some customers ordering several systems at once. Watch out for details in future editions of the journal.

I hope you enjoy the issue, Dr David Martyr CEO

*Product coming soon. **MAPlinx is currently available for internal development only; for external development coming soon.

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TECAN JOURNAL 2/2018

CONTENTS

Contents 2

CEO Welcome

4 - 5 Putting stem cells at the heart of predictive drug toxicity testing

6 - 7 Is NGS costing you more than you think?

8-9 Building blocks for custom solutions

8 - 9

Building blocks for custom solutions

10 - 11

Redesigning the drug safety workflow

12 - 13

Sequencing made easy

14 - 15 Automation by numbers 16 - 17 Cell counting made easy 18 - 19

Long-read sequencing for improved analysis

20 - 21 Lipidomics – the future of personalized medicine?

22 - 23

Biosensors light the way for drug development

24 - 25 A tailored solution to meet bioprocessing challenges

28 - 29 Long-term partners in success

26 - 27 Making big discoveries accessible to all laboratories

28 - 29 Long-term partners in success 30 - 31 Detection at the molecular level

Applications and platforms presented in the Tecan Journal may not be available in all markets. Please consult your local Tecan office for information.

TECAN JOURNAL 2/2018

3

DRUG DISCOVERY

Putting stem cells at the heart of predictive drug toxicity testing Failure of candidate drugs in late-stage clinical development continues to be a challenging and costly problem for the pharmaceutical industry. A recent study of 640 novel therapeutics found that more than half (54 %) failed in clinical development, most (57 %) due to insufficient efficacy, and a sizable proportion (17 %) due to safety concerns.1 The earlier in the drug development process that these issues can be accurately assessed, the lower the risks – both financially and to human health – with taking a compound forward into clinical testing.

Stem cells are playing a growing role

electrophysiological changes that can

The protocols required to differentiate

in all phases of drug discovery – from

adversely affect myocardial contractility.

stem cells into the intended cell type

disease modeling to the development

In a comprehensive review article,

are equally complex, requiring close

of innovative therapies – providing

authors from the US Food and Drug

monitoring and the addition of precise

self-renewing, sustainable sources of

Administration discuss the advantages

amounts of various growth factors and

differentiated cell models. Derived

of using hiPSC-derived cardiomyocytes

other biological and chemical stimulants

from either human embryonic stem

to assess drug-induced structural

at key time points. Any inconsistency in

cells (hESCs) or induced pluripotent

cardiotoxicity.5 This article emphasizes

how these protocols are carried out – eg.

stem cells (hiPSCs), these models are

the renewable and readily scalable nature

due to variability between the technique

becoming increasingly popular for in

of stem cells, and the benefits of testing

of technicians – can affect the behavior

vitro preclinical predictive toxicology

directly in human cardiomyocytes rather

of the resulting cardiomyocytes in

testing, providing more physiologically

than having to translate findings from

subsequent toxicity testing.

relevant data to detect potentially

toxicity studies conducted in animal

harmful on- and off-target effects

models or animal cell lines. It concludes

Automated liquid handling and cell

before drug candidates reach the

that high throughput screening using

processing technologies can minimize

clinical phase.2,3 This helps to reduce

a hiPSC-cardiomyocyte model is a

this variability, as well as eliminating

attrition rates, costly program failures

‘feasible approach to assess potential

much of the tedious, time-consuming,

and post-market withdrawals, and can

contractile and structural cardiotoxicity

and labor- and cost-intensive aspects

even provide the opportunity for cost-

in early phase drug development’, and

of working with stem cells and stem

efficient modification of the drug to

supports combining structural and

cell-derived models. These benefits

minimize its toxicity.

electrophysiological tests on the same

become even greater as methods are

platform.

scaled up to meet the demands of large-

Using stem cells to detect drug

scale applications. High throughput

cardiotoxicity earlier

The challenges of working with

automation solutions are essential in

Cardiotoxicity is one of the top

stem cells

enabling stem cell-derived models, such

three causes of post-marketing

Stem cells are highly sensitive to

as hiPSC-derived cardiomyocytes, to be

drug withdrawals due to adverse

variability in their environment and

applied as a pharma industry standard

reactions. Rapid advances are being

handling, and can be tedious and tricky

for earlier predictive toxicity testing in

made in the use of hiPSC-derived

to work with. Nurturing stem cells to

drug discovery.

cardiomyocytes to test drug candidates

grow and reproduce in culture is as

for their potential to cause structural or

much art as science.

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The Blog 4

TRENDS, NEWS, STORIES AND MUCH MORE! FROM THE EXPERTS TO YOU.

TECAN JOURNAL 2/2018

DRUG DISCOVERY

References 1) Hwang, TJ et al. Failure of investigational drugs in latestage clinical development and publication of trial results. JAMA Intern Med, 2016, 176(12), 1826-1833.

2) MacDonald, A. Stem cells in drug discovery. Technology Networks, April 4, 2017. https:// www.technologynetworks.com/cellscience/articles/stem-cells-in-drugdiscovery-286825 3) iPS Cell Drug Discovery Taking Off with First Clinical Trial. Press release. August 2, 2017. https://asia. nikkei.com/Tech-Science/Science/iPScell-drug-discovery-taking-off-withfirst-clinical-trial

If you haven’t listened to it already, check out the recent webinar entitled Automated assessment of liver and cardiac toxicities in lead optimization, using biochemical and human iPS cell assays on the Tecan FluentŽ at www.drugtargetreview.com/ webinar/30994/automated-assessmentof-liver-and-cardiac-toxicities-in-leadoptimisation-using-biochemical-andhuman-ips-cell-assays/

4) Onakpoya, IJ et al. Post-marketing withdrawal of 462 medicinal products because of adverse drug reactions: a systematic review of the world literature. BMC Med, 2016, 14, 10.

5) Yan, X and Papoian, T. Moving beyond the comprehensive in vitro proarrhythmia assay: use of humaninduced pluripotent stem cellderived cardiomyocytes to assess contractile effects associated with drug-induced structural cardiotoxicity. J Appl Toxicol, 2018, Feb 27. doi: 10.1002/jat.3611. (Epub ahead of print)

www.tecan.com/blog TECAN JOURNAL 2/2018

5

GENOMICS

Is NGS costing you more than you think?

The sequencing, analysis and data storage costs associated with NGS are dropping rapidly, making this powerful technology ever-more affordable and widely accessible. Cost analyses indicate that the landmark $100 genome is almost within reach, but the reality is that most NGS labs are still spending far more than that. Here we explore some hidden costs of NGS that occur upstream of sequencing, highlighting some of the ways labs are hemorrhaging money unnecessarily, without even realizing it.

Looking upstream of sequencing

Nucleic acid extraction – The ‘massively

or add something twice. At best, such

There have recently been several

parallel’ high throughput nature of

mistakes can mean that the process

articles looking at the ‘real cost of NGS’,

NGS inevitably highlights any upstream

needs to be repeated. At worst, errors

but most of these focus on everything

bottlenecks in sample preparation, often

such as cross-contamination can go

from sequencing onwards, and gloss

in nucleic acid extraction. Conventional

undetected, leading to erroneous results

over the upstream steps of sample and

DNA and RNA isolation methods are

and potentially serious consequences,

library preparation. Unfortunately, that

both low throughput and very labor

especially in a clinical setting.

is a significant oversight, as the NGS

intensive, decreasing productivity.

process starts long before you press

Manual extraction approaches also

PCR for library preparation and

‘go’ on your sequencer. The good news

increase the risk of inaccuracy and

quantitation – It is worth paying special

is that many of these ‘hidden’ costs can

human error, which ultimately translate

attention to PCR workflows that are

be easily reduced and controlled, if you

into wasted time, reagents and

integral to library preparation and

know where to look. The main culprits

resources. And, once extracted, nucleic

quantitation, as cross-contamination

in NGS are:

acids need to be accurately quantified

can be a major issue that is hard to

for subsequent titration. Using robotics

detect. The nature of PCR means that

to work with the smallest volumes

small errors are readily amplified and

misplace or mix up samples in a high

possible at this stage can significantly

can skew your library. For example,

throughput NGS workflow, leading

lower the price per sample.

pipetting the wrong index into the

Tracking sampling errors – It is easy to

wrong sample could lead to hugely

to re-collection or repeat testing. In some cases, replacing samples may

Library preparation – If you have

significant knock-on consequences.

not even be an option, or a mix-up may

large numbers of samples to process,

This can be particularly problematic

go undetected. Occasional errors like

library preparation can actually cost

when the aim is to assess rare

this may not seem like a big deal, but

more than the sequencing itself. With

variants (eg. when examining tumor

tracking down lost samples and re-

many steps involved for amplification,

heterogeneity). And, when PCR is used

running tests is a drain on resources,

pooling, normalization, etc., there are

for quantitation, everything is typically

and can add up to serious money. In

numerous opportunities for waste and

done in triplicate, multiplying the total

addition to the direct costs, there is

error. It is critical that reagents, samples

number of pipetting steps and different

also the incalculable impact on clients

and controls are added in the right

samples you have to deal with, further

and patients, ranging from simple

amounts, to the right wells, at the right

increasing the risk of errors. It is difficult

dissatisfaction to poor medical decisions

time. Since the volumes involved are

due to lack of information or an

very small – only a few microliters – it

erroneous result. End-to-end traceability

is easy to accidentally skip a sample

is therefore vital at every step of the NGS workflow, from sample collection right through to reporting.

The Blog 6

TRENDS, NEWS, STORIES AND MUCH MORE! FROM THE EXPERTS TO YOU.

TECAN JOURNAL 2/2018

GENOMICS

to estimate the frequency and ultimate

Check out our dedicated genomics

cost of these sorts of mistakes, as many

channel – The Helicase – at

of them may go undetected, so it is

www.tecan.com/helicase for

better to take measures to avoid them in

additional information and support on

the first place.

automating your genomics workflows.

Understanding the real cost of NGS This list is far from exhaustive, but hopefully the issues discussed here make it clear that taking time to scrutinize the NGS process from start to finish – especially the steps upstream of sequencing – is a worthwhile endeavor.

Investing in quality automation can go a long way to addressing all of the above issues by eliminating unnecessary manual steps and sources of error. At the same time, it can free up skilled staff for more important (and interesting) work.

www.tecan.com/blog TECAN JOURNAL 2/2018

7

OEM SOLUTIONS

Building blocks for custom solutions Developing customized laboratory automation solutions for complex liquid handling operations requires access to state-of-the-art robotics and OEM instrumentation. Flexible and modular components are essential building blocks of many of these bespoke designs, and the Xantus® XYZ robot has stood the test of time, supporting over 10 years of OEM integration by Synchron Lab Automation.

Synchron Lab Automation – based in

company became a Tecan distributor

“Customers turn to us when a standard

Etten-Leur, the Netherlands – designs

around 1980, and my father – who also

pipetting platform does not meet their

and builds software and hardware for

worked in the business – was

requirements. Every client has an idea of

automated liquid handling systems,

subsequently asked to develop some

what they want, but we help to make it a

customized to its clients’ specifications

ELISA software for the Tecan MiniPrep,

reality, developing software and

and needs. The company has a long-

and this was the beginning of Synchron.

hardware and rapidly 3D printing

standing history of working with Tecan,

When Tecan decided to invest in direct

prototypes, prior to creating the final

as Derk Wilten, Director and Owner,

sales to its European customers, we

design. Our expertise in custom lab

explained: “Synchron was born out of

partnered with Sias, and now the story

automation has led to us working with

the automation department of Wilten

has come full circle; my father worked

customers in Europe, Singapore and the

Instruments, a laboratory supplier

with Tecan, then Sias, and now Sias is

USA, on projects ranging from benchtop

started by my grandfather in 1952. The

Tecan again.”

automation to more complex systems,

Derk Wilten, Director of Synchron Lab Automation

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TECAN JOURNAL 2/2018

OEM SOLUTIONS

The Xantus robot’s open geometry, modularity and flexibility have made it the best solution on the market for integration into our projects.

The Xantus robot offers a wide range of integration options

all dependent on our specialized

channels, with or without a gripper –

need additional training, we know where

knowledge of liquid handling robotics.

and we can even mount the arm to a

to turn. Tecan is a great company to

Rather than limiting ourselves to a single

wall, removing the deck entirely.”

partner with, and there’s no doubt that

market, we recognize that our expertise

the Xantus robot will continue to be a

can be applied across the board. Our

“The Flexispan feature – enabling

central feature in many of our future

customers therefore come from a full

individual channels to move

projects,” Derk concluded.

range of industries, including life

independently along the Y- and Z-axes

sciences, petrochemistry, pharma, food

– is also highly valued, and each channel

and beverage, and agriculture.”

can be fitted to a range of pumps and use either disposable or fixed tips. The

To find out more about partnering with Tecan, visit

“Although every project is different, they

new micropump technology also gives

all share some similarities, with liquid

us a competitive advantage, by allowing

handling at the heart of each design. If

us to accurately dispense both large and

we develop a technique to pipette milk

small volumes using the same pump.

To learn more about Synchron Lab

for a dairy, for example, the same

Our customers expect reliability and the

Automation, go to

technology can serve a similar need in a

system needs to be the best of the best.

www.synchronlab.com

blood bank. We look to build as much

The Xantus robot is extremely high

standardization as possible into our

quality and reliable. The precision

designs, and so it is essential to have

mechanics in the robotic arm are a feast

access to as many ‘building blocks’ as

for the eyes but, more importantly, they

possible, which is where the Xantus

are very easy to access for servicing –

robot comes into play. Its open

it’s a fine piece of Swiss engineering.”

partnering.tecan.com

geometry, modularity and flexibility have made it the best solution on the market

“We have now been working with the

for integration into our projects; 95 % of

Xantus robot for over 10 years, so we

our projects make use of this platform.

know exactly what configuration to buy

The configuration of the system is

and what to do with it. However, we still

amazing. We can get it in any size up to

meet with the Tecan team twice a year

three and a half meters long – with one

to discuss our projects and, if we ever

or two arms, up to eight pipetting

TECAN JOURNAL 2/2018

9

DRUG DISCOVERY

Redesigning the drug safety workflow Detecting adverse off-target

Safety pharmacology is an important

a-chip approaches – that enable drug

effects is crucial to ensure the

part of the drug discovery process,

safety studies to be performed far earlier

safety of potential therapeutics,

identifying and investigating any

in the development process. Ncardia is

potential undesirable pharmacological

a pioneer in this area, developing and

effects of candidate drugs before they

commercializing highly predictive cellular

enter clinical development. These effects

assay systems derived from human stem

have traditionally forced

are often hard to characterize in cellular

cells to accurately assess drug safety.

pharmaceutical companies to

or simple in vivo models, requiring

Dr Stefan Braam, Co-founder and CEO

perform safety pharmacology

animal testing – usually in canine models

of Ncardia, explained: “Our aim is to

studies at a late stage of the

– to ensure the potential therapeutic

put ‘human’ testing at the forefront

is safe to proceed to the next phase.

of drug discovery, by developing the

The cost and ethical issues associated

best possible human cellular models.

with such testing mean that safety

This is very attractive from an industry

models and automated screening

pharmacology studies have traditionally

perspective, as it ensures only viable

processes are revolutionizing

been performed at a late stage of

candidate drugs are progressed through

drug safety studies, enabling

preclinical development. This can lead

the pipeline, while avoiding the risks

much earlier testing, with

to late-stage failure of candidate drugs,

and ethical considerations associated

resulting in significant financial losses.

with animal testing. Our expertise in the

but limited throughput and ethical considerations

drug development process.

Human stem cell-based cellular

companies such as Ncardia at

manipulation and differentiation of stem

the forefront of this workflow

Avoiding these expensive failures

cells into cardiac cell lines allows us to

transformation.

is a major goal for pharmaceutical

create a synchronously beating layer

companies, creating a real need for

of human cardiomyocytes in culture. In

new technologies – such as organ-on-

combination with predictive safety tests

Stefan Braam (left) and Farbod Famili with Ncardia’s Fluent Automation Workstation

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TECAN JOURNAL 2/2018

DRUG DISCOVERY

– such as multi-electrode arrays – these

applications. We therefore wanted

you the option to define everything –

cells provide a convenient and cost-

to automate the entire cell culturing

labware, protocol availability, dispense

effective cardiac model, allowing safety

process, performing each step as quickly

volumes, etc. – for inexperienced

studies to be performed much earlier

as possible to avoid variations across

operators, which makes it much easier

in the overall process. Our technology

a microplate. We looked at the various

for everyone in the lab to use the system

has already been adopted by many

liquid handling solutions on the market,

once it is in routine use. Working in

of the world’s top 20 pharmaceutical

and were impressed by both the speed

384-well microplates, we’re currently

companies, demonstrating the huge

and flexibility of the Fluent® Automation

handling 40 plates in a single run. This

potential of this approach.”

Workstation. With three independent

would take at least five times as long

arms, it offered us the potential to

to do manually, so the benefits are

Assay development is at the forefront

significantly accelerate the process, while

felt immediately.”

of Ncardia’s offering, and the company

still being a very versatile platform. We

is actively involved in the design and

had already had a positive experience

validation of assays intended for

working with Tecan when we purchased

day-to-day use by pharmaceutical

a Spark® microplate reader, and so we

and biotechnology companies. Stefan

were confident in both the service and

continued: “Safety pharmacology is just

support the company offers.”

one potential application of human-

For more information on Tecan’s drug discovery portfolio, visit www.tecan.com/drugdiscovery

derived cellular models, and we are also

“The system was installed at the

interested in using this technology to

beginning of summer 2017, and has

To learn more about Ncardia,

enable high throughput drug efficacy

everything required to culture and

go to www.ncardia.com

testing. By culturing cardiomyocytes in

differentiate stem cells integrated into

a 384-well microplate format, we can

it, including an automation-friendly

provide our customers with the tools to

incubator and a laminar flow HEPA

screen tens of thousands of potential

hood to ensure sterility. We knew that

drug molecules in a human model. This

setting up such a complex workflow

ensures more reliable and accurate

for highly sensitive cells would be

screening data, reducing the risk of

challenging, and finding the optimal

throwing out the baby with the

labware and workdeck configuration was

bath water.”

the first obstacle, but the 3D simulator in FluentControl™ makes it very easy

Farbod Famili, Assay Development

to visualize your protocols, speeding

Scientist at Ncardia’s Leiden facility

up process development. It also gives

in the Netherlands, took up the story: “Pharmaceutical companies understandably demand assays that are robust and reliable, and compatible with high throughput automated screening platforms. When I first arrived at Ncardia, almost all of our processes were performed manually, which worked when handling a limited number of compounds for safety pharmacology

We had already had a positive experience working with Tecan… so we were confident in both the service and support the company offers.

studies using multi-well, multi-electrode array chips, but was not compatible with high throughput efficacy testing

TECAN JOURNAL 2/2018

11

GENOMICS

Sequencing made easy Automation is helping the University Hospital Münster to streamline its molecular diagnostics workflows, increasing throughput for Sanger sequencing and NGS library preparation.

The University Hospital Münster (UKM)

workflows, as Dr Hartmut Schmidt, Head

Hartmut continued: “One of the major

is one of Germany’s leading hospitals,

of Molecular Diagnostics, explained:

challenges in Sanger sequencing is

providing state-of-the-art healthcare to

“One of our department’s main roles is

the removal of unincorporated dye

national and international patients. It is

to perform routine Sanger sequencing

terminators. In the past, we did this

also home to the Center for Laboratory

on samples from several different

manually, but it was a slow process that

Medicine, which provides core

groups. We can carry out as many as

required three hours of preparation

laboratory services – including clinical

2,000 analyses per day, occasionally

before sequencing could begin. Not

chemistry, hematology, endocrinology,

even more; the human genetics

only was this time consuming, but it

virology, immunology and therapeutic

department, for example, sometimes

restricted our flexibility in the lab, and so

drug monitoring – to the hospital and

submits nine plates for processing on

we turned to automation.”

its external clients, typically performing

one day, which is around 900 samples

around 10,000 analyses per day. The

from just one group.”

laboratory’s molecular diagnostics department uses Sanger sequencing with capillary electrophoresis for routine diagnostics, as well as performing next generation sequencing (NGS) in collaboration with other UKM groups.

Automation is the key to standardizing and streamlining the department’s

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TECAN JOURNAL 2/2018

One of the big advantages of automating the workflow is the elimination of dye blobs… enabling faster analysis of the data.

GENOMICS

Left to right: Mathias Isakiewitsch, Hartmut Schmidt, Kathrin Meyer zu Himmern and Kathrin Haar with the Freedom EVO workstation

The laboratory chose a Freedom EVO®

to medium to high; if the signal is too

chemistries just by using different

150 workstation equipped with a

high, the analysis may fail and have to

automated methods; it’s a really good

MultiChannel Arm™ 96 to help with

be repeated. We use a magnetic bead-

system, and one that can evolve to

the dye removal. This set-up allows

based method of purification, and have

accommodate other chemistries in the

one plate to be incubated while the

been able to implement a signal cut-off

future as our needs change.”

workstation performs the necessary

value by using the Freedom EVO to

pipetting steps to prepare the next,

add a defined amount of beads. This

In a busy department, staff only have a

which is an important feature to meet

allows us to remove any unincorporated

limited amount of time to spend learning

the lab’s testing demands. “Automation

dye terminators, maintaining the signal

a new technique, and one of the big

of the clean-up procedure has improved

intensity within a suitable range and

advantages of the Tecan systems for

our throughput – we can do up to six

reducing the number of repeat sample

UKM is that they are easy to use. The

runs in one hour – allowing us to handle

analyses, something we were unable to

company’s service engineer helped UKM

a greater number of samples and report

do with our previous filtration-based

staff to write TouchTools™ protocols that

the results more quickly,” Hartmut

purification process.”

take operators through the workflows

commented.

step by step, using photographs integrated In addition to Sanger sequencing, the

into each program to display on screen

“As an accredited laboratory, we take

molecular diagnostics department

exactly where the labware, samples

great care to ensure the quality and

performs automated NGS library

and reagents should be placed on the

reliability of our sequencing results,

preparation for a number of other

workdeck. “It is a really useful feature that

and thoroughly compared our old

UKM groups. This work is performed

makes the platform very straightforward to

method with the automated protocol

on a Freedom EVO NGS workstation,

use,” Hartmut concluded.

before it was implemented. One of

which is available with a number of

the big advantages of automating

verified protocols – developed in

the workflow is the elimination of dye

collaboration with the vendors – for

To find out more about Tecan’s

blobs – artefacts that interfere with the

different chemistries. Tecan application

genomics solutions, visit

sequencing analysis, making it harder to

specialists worked with the UKM team

www.tecan.com/genomics

identify the correct base pairs – enabling

to implement and evaluate these

faster analysis of the data. We have

protocols for the chemistries currently

also been able to overcome the issue

used by the human genetics and

of variations in signal strength. The

neurogenetics departments. Hartmut

samples sent to us for analysis by other

added: “Now that the evaluation is

groups vary in signal intensity from low

complete, we can process these varying

To learn more about UKM’s Center for Laboratory Medicine, go to www.ukm.de

TECAN JOURNAL 2/2018

13

BIOPHARMA

Automation by numbers Biopharmaceutical production is a complex process compared to small molecule drug manufacture, requiring the interaction of multiple biosynthesis pathways to create the target product. KBI Biopharma is using high throughput bioanalysis to gain an in-depth understanding of these biological processes, ensuring efficient, reproducible and scalable biomanufacturing processes.

Carolina, and works with a wide range of

analyst – which we were able to achieve

clients – from biotech start-ups to big

and validate within six weeks of

pharma – to accelerate and optimize

installation. This automated version of

their drug development. The company’s

the assay offered at least comparable

highly successful biological

performance to our manual assay, and

manufacturing programs rely on using

took the same amount of time to run

advanced analytical techniques to

– 3.5 hours. We then looked at how to

acquire in-depth knowledge of each

use the instrument’s worktable capacity

biosynthesis pathway, resulting in more

and scheduling capabilities to save time

robust and scalable processes.

by processing multiple assay plates in parallel, something that would be

This data-driven approach requires large

virtually impossible for a single analyst

numbers of samples to be tested for

to achieve manually.”

each new project, and the company’s analytical development team recently

Devon continued: “The first step for any

Members of the analytical development team

began investing in automated liquid

parallel assay is to work out the on-deck

(left to right): Kirsten Dysinger, Elijah Gaspard,

handling solutions to enhance its high

capacity of every element required to run

Devon Davenport, Phoebe Lee and Derek Ryan

throughput testing capabilities. Devon

the assay – including hardware modules,

Davenport, Scientist I, explained: “When I

assay plates, labware, consumables and

arrived at KBI in 2014, there was no

reagents – then back-calculate the

automation in use for our ‘platform’

maximum throughput based on the

assays; everything was performed

limiting factor. For the HCP assay, the

Biopharmaceutical production requires a

manually. Coming from a high

volume of the reagent trough determined

wide range of studies – including

throughput automation background, I

that we could run eight plates without

immunoassays, enzymatic assays,

could immediately see the benefits of

the need for user intervention, but there

residual DNA detection and

moving some of our high volume residual

were only six assay positions on the

chromatography – to fully understand

assays to an automated platform, and

worktable. We therefore built a script

and optimize the biomanufacturing

started to explore the options available.

around parallel processing of six assay

process. Many of these analytical

My manager, Alan, and I each already had

plates. This protocol took just under five

activities are both time consuming and

some experience with Tecan platforms

hours to run all six plates – a huge time

complex, leading many

from previous roles and so, in July 2016,

saving – but still resulted in instrument

biopharmaceutical manufacturing

KBI purchased a Freedom EVO® 200 to

‘idle time’ and unused reagent capacity,

facilities to use automated or semi-

automate our HCP ELISAs. This one

and so we ‘looped’ two additional assay

automated workflows. This not only helps

assay was occupying four members of

plates onto the end of the protocol by

to increase throughput and improve

staff full time, and our intention was to

exchanging the completed plates with

consistency, it also frees up technicians

offload all of this work onto the

two fresh ones from the instrument’s

to perform tasks that are less automation

automated platform, allowing the

labware hotel. As a result, we can now

friendly or to analyze results.

analysts to perform other activities.”

process eight complete assay plates in 9

KBI Biopharma is a contract development

“We began by setting up a ‘single plate’

for user intervention, compared to 28

and manufacturing organization

assay protocol on the Freedom EVO –

hours of hands-on analyst time. We have

headquartered in Durham, North

effectively taking the place of one

since applied this same approach to a

hours and 37 minutes without the need

14

TECAN JOURNAL 2/2018

BIOPHARMA

The systems’ Freedom EVOware Plus has been great to work with; it’s very user friendly and intuitive.

number of other HCP ELISA protocols – each designed to meet individual clients’ specifications – with similar results.”

“Following the success of the HCP project, we have now also begun automating our residual DNA platform method using a Freedom EVO 100, based on the positive experience we’d had with the HCP platform and the ease of scripting within the software. The systems’ Freedom EVOware® Plus has been great to work with; it’s very user friendly and intuitive, and offers advanced users the ability to change and modify far more parameters than most other laboratory automation software I’ve worked with. This allows you to edit, for example, the exact timings and durations of individual pipetting steps or labware movements, which proved critical to achieving an optimized six-plate HCP assay. Building on this, we have now created modular automation ‘nodes’ which we can link together to form a complete protocol using the different, client-specific HCP assays running in parallel. The local Tecan team – who happen to be just up the road – have been great to work with, and have been extremely supportive as I’ve tried to get the most out of the system.

Introducing the Tecan platforms has been so successful that we’re considering an

To find out more about Tecan’s

additional Freedom EVO 200 to provide

Freedom EVO platform, visit

extra capacity and allow more assay development,” Devon concluded.

www.tecan.com/freedomevo To learn more about KBI Biopharma, go to www.kbibiopharma.com

TECAN JOURNAL 2/2018

15

CELL IMAGING

Cell counting made easy Small molecule drug discovery involves a range of functional assays that have traditionally relied on manual cell counting techniques to monitor proliferation, migration and invasion.

Automated cell counting is enabling the EB House Austria to save time and free up personnel, as well as designing time-course experiments that were previously unachievable.

The EB House Austria is a unique,

while the gene therapy team is hoping

entered phase III clinical trials. Our

highly specialized clinic for patients

to tackle the problem at its root, by

attention then turned towards treating

with epidermolysis bullosa (EB), a rare

replacing or correcting genes using

RDEB, where we are currently studying

inherited disease that causes the skin to

different technologies, such as

differently regulated microRNAs in

become ‘as fragile as a butterfly’s

CRISPR, conventional DNA therapy or

developing tumors to identify potential

wings’, blistering in response to friction

trans-splicing.”

biomarkers.”

or trauma. The clinic, located within the University Hospital of Salzburg, was

“Our research covers the full spectrum

“A large variety of functional assays

the initiative of patient organization

of the disease, from the milder EB

were needed to support our research,

DEBRA Austria, and opened in 2005.

simplex (EBS) to the most severe form

and we purchased a Spark® multimode

The EB House consists of three units:

– recessive dystrophic epidermolysis

microplate reader back in 2016 to meet

the EB outpatient clinic; the EB

bullosa (RDEB) – which ultimately leads

the workload. We carry out confluence

academy; and a research arm

to premature death from chronic

measurements for many proliferation,

developing therapies to treat the

wounds that develop into cancers.

migration and invasion assays,

disease. The clinic’s 30-strong group

Our initial focus was on EBS, where

as well as using luminescence to

of researchers is divided across three

we discovered an inflammatory pathway

study the interactions of microRNAs

areas: gene therapy; basic research into

that is constitutively activated in the

with three prime untranslated regions

cancer and wound healing; and small

patient population. We repurposed the

(3’-UTRs), and to assess cell viability

molecule development and epigenetics.

interleukin-1 beta inhibitor diacerein –

with tetrazolium dye MTT assays.

Dr Verena Wally, Group Leader of

which is used in the treatment of

We also investigate transfection

Small Molecules Research, explained:

osteoarthritis – to treat these patients.

efficiencies, for example by

“The main aim of my group is to

This project has since been out-licensed

looking for green fluorescent

identify potential therapeutic targets,

to a pharma company, and has now

protein expression.”

Members of the EB House research team (left to right): Verena Wally, Michael Ablinger, Thomas Lettner, Roland Zauner, Monika Wimmer and Melanie Böhm

16

TECAN JOURNAL 2/2018

CELL IMAGING

eproducibility R has been a key advantage of the new platform, as the reader removes any user bias or human error, and we can now measure even smaller cell sizes.

“In September 2017, we purchased the

scale migration assays involving 12

“We chose to purchase the Spark on the

Gas Control Module (GCM™) and Live

plates, something we couldn’t do

back of a conversation with another lab,

Viewer, which offered us the expanded

previously. Ultimately, the greatest

and Tecan’s reputation for excellent

functionality we need. The reader has

benefits have been the time saved and

customer service. We have been

brought huge benefits to our workflow.

the personnel it frees up. The platform

impressed by the responsiveness and

Previously, we were carrying out

now runs every day, and is used by all

level of technical support that we have

migration assays by hand; taking

the groups.”

received. We’re fortunate to be only 20 kilometers from the Tecan office and, if

pictures with a microscope and using cell counting software or counting them

“We are currently preparing to run

we have a problem, a Tecan team

ourselves. For luminescence assays, we

time-course experiments over 24 hours,

member gives us a call or visits the site;

were fortunate to be able to use

measuring every two hours to monitor

it was a deciding factor in choosing to

systems from other departments, but it

the proliferation and viability among

upgrade the system,” Verena concluded.

was time consuming to move between

other factors. Before the Spark,

locations, particularly when running

we were limited to taking

time-course experiments.

measurements of time points when

Reproducibility has been a key

we were actually in the lab – say after

To find out more about live cell

advantage of the new platform, as the

eight hours, 12 hours and 24 hours –

imaging, visit

reader removes any user bias or human

but this could lead to us missing

error, and we can now measure even

the time point that would actually be

smaller cell sizes, expanding the range

important. Leaving the plate in the

of cell types we can analyze. In

reader and taking readings every two

addition, the Spark can handle higher

hours will be hugely beneficial. Gone

density plates, enabling us to measure a

are the days when if we missed the

House Austria, go to

greater number of samples in a single

event, we had to start again from

www.eb-haus.org/en/home.html

run. We can also now carry out large-

the beginning.”

lifesciences.tecan.com/live-cellimaging To learn more about the EB

TECAN JOURNAL 2/2018

17

GENOMICS

Long-read sequencing for improved analysis Next generation sequencing is now in widespread use throughout the life sciences sector, but the commonly used short-read sequencing methods are often subject to GC base pair bias. Combined with the inherent mapping ambiguity of the short reads, this often results in fragmented genome assemblies, creating a demand for technologies offering longer reads that simplify analysis and yield more complete sequences. Using its proprietary technology, Pacific Biosciences is able to offer longer reads, more uniform coverage and high accuracy, supporting advanced genomics, full-length transcript sequencing and epigenetics.

“The major advantages of our technology are the long reads, uniform coverage and high consensus accuracy. This accuracy is due to low sequencing-context bias, very low systemic error and the random nature of read errors. For instance, with 10- to 20-fold coverage we can achieve 99.9 % accurate reads, which can be readily increased to 99.999 % accuracy with additional sequencing coverage.

Our system can also accommodate a broad range of template sizes – from ~200 to >10,000 bp – making it suitable for a wide variety of applications, including whole genome and targeted sequencing. As a result, SMRT Sequencing technology is proving popular for plant and animal sciences research – particularly in academic labs – as well as in the human biomedical space, for applications

The Cavro ADP is at the heart of the Sequel System

such as HLA typing.”

Pacific Biosciences (PacBio) is a

(ZMWs), which you can think of as very

sequencing technology provider based

small reaction vessels. We anchor the

“Our original system – the PacBio RS II –

in Menlo Park, California. Using its

polymerase with the pre-bound DNA

had 150,000 ZMWs per SMRT Cell,

proprietary Single Molecule, Real-Time

template at the bottom of the ZMW,

equating to about 55,000 reads, and

(SMRT®) Sequencing technology, the

and introduce fluorescently-tagged

could process up to 16 cells per run. This

company has developed two long-read

nucleotides. The ZMW is illuminated

may sound like a very large number, but

sequencing systems – the PacBio RS

from below, and the attenuated light

throughput was still a limitation, so we

II and the Sequel® System – offering

enters the lower 20-30 nm of each

launched the Sequel System in October

a comprehensive view of genomes,

ZMW, exciting the fluorophore as the

2015. With this platform, each SMRT Cell

transcriptomes and epigenomes.

polymerase incorporates each base

contains one million ZMWs, offering a

Marty Badgett, Senior Director of

into the native strand. Our instrument

sevenfold improvement in throughput,

Product Management at PacBio,

basically carries out signal processing

and we’re currently working on a higher

explained: “Our instruments sequence

by ‘eavesdropping’ on the process of

density SMRT Cell containing eight

DNA directly in SMRT Cells packed with

DNA synthesis as it happens.”

million ZMWs.”

thousands of zero-mode waveguides

18

TECAN JOURNAL 2/2018

GENOMICS

Automation of NGS protocols is now crucial for many life sciences applications

Paz Yogev, a Staff Mechanical Design Engineer at PacBio, added: “When we were developing the Sequel System, we wanted to incorporate a number of additional liquid handling features that the pipettor used in the PacBio RS II simply couldn’t offer. We researched a

his OEM solution offers a combination T of performance, accuracy, cost and advanced features… that is very favorable for our application.

number of off-the-shelf solutions – as well as the possibility of developing a customized pipetting system based on one of these options – before eventually deciding on Tecan’s Cavro

®

Air Displacement Pipettor (ADP). This OEM solution offers a combination of performance, accuracy, cost and advanced features – including automatic tip detection and ejection, plus liquid level detection – that is very favorable for our application. We initially bought a couple of sample units and set up a test rig to run comparisons. This was both easy and inexpensive, making development of the pipetting system

same supplier maximizes the likelihood of everything working in synergy

To find out more about

without any issues.”

partnering with Tecan, visit partnering.tecan.com

“We have been working with the Cavro ADP for several years now, and it has been really good to us – it’s very reliable. Our field service engineers respond to any problems encountered

To learn more about Pacific Biosciences, go to www.pacb.com

by customers, and we very rarely have to reach out to the Tecan team for extra support, but we know it’s there if we need it,” Paz concluded.

very cost effective. In addition, we chose to use Tecan’s disposable tips, as buying the pipettor and consumables from the

TECAN JOURNAL 2/2018

19

MOLECULAR DIAGNOSTICS

Lipidomics – the future of personalized medicine?

Personalized medicine is on the horizon, and cell membrane lipidomics may hold the key. Italian biotechnology company Lipinutragen is studying the relationship between the membrane status and dietary and health conditions. A crucial part of the process is the isolation of mature red blood cells, which can be performed more precisely and reproducibly using laboratory automation than by manual processing. of a balanced diet. The main cause of a nutritional imbalance is ignorance. People know that a lack of fruit in the diet can lead to a vitamin deficiency, but are simply not aware of the need to consume omega-3 and -6 fatty acids, which can lead to the development of essential fatty acid deficiency syndrome.”

“Sample preparation for lipidomic analysis is a laborious, time-consuming, multi-step process that is difficult to perform precisely by hand, and so we automated the workflow on a Freedom EVO® platform. Blood samples are collected in tubes containing EDTA – the only anticoagulant able to The Lipinutragen team (left to right): Andreea Popa, Carla Ferreri, Anna Vita Larocca, Simone Deplano, Valentina Sunda and Chryssostomos Chatgilialoglu

sequester calcium – and delivered to the laboratory within 24 hours. After centrifuging, plasma, platelets and lymphocytes are removed leaving the

Bologna-based Lipinutragen, a spin-out

and specialist pharmacies, which

red blood cell layer. Red blood cells have

of the Italian National Research Council

can offer advice about dietary

a four-month life span, and the mature

(CNR), was founded in 2005 by

changes and supplements to address

cells that we require are smaller in

Drs Chryssostomos Chatgilialoglu

a lipid imbalance.”

diameter and heavier than the others. We

and Carla Ferreri, who drew on their

measure the diameter of the red blood

expertise in synthetic and analytical

Carla continued: “Typically, the first

cells using a SCEPTER™ 2.0 handheld

chemistry to establish a lipidomics

sign of a lipid imbalance is stress.

cell counter (EMD Millipore), selecting

laboratory, focusing on the isolation

Biochemical analysis may not give

those that are six microns across.

of mature red blood cells to allow

any indication of a pathology, but

These cells are removed and placed in

analysis of the membrane fatty acid

the person’s mood may change, or

water to rupture the membrane, and

profile. Carla, now Lipinutragen’s R&D

they might not sleep well and still

then centrifuged to produce a pellet.

Director and a Senior Researcher at

feel tired when they wake up in the

The phospholipids are extracted from

CNR, explained: “The lipid profile of the

morning. These are all signs of a general

the pellet using chloroform:methanol

mature red blood cell membrane is a

imbalance, including the cell membrane

(2:1, v/v) and derivatized to form fatty

comprehensive biomarker of a person’s

lipid profile. The membrane contains

acid methyl esters for analysis by gas

state of health, quality of nutrition and

two types of fats; those that can be

chromatography. All stages except

lifestyle. This has benefits in both clinical

produced by the body’s enzyme system,

centrifugation, evaporation and GC

applications – enabling the development

and those such as omega-3 and -6 fatty

analysis have been automated, which has

of personalized treatment strategies –

acids, which must be consumed as part

made a big difference to our laboratory.”

20

TECAN JOURNAL 2/2018

MOLECULAR DIAGNOSTICS

The result was a liquid handling system that is far more consistent and precise than manual pipetting, which is exactly what we need for lipidomic analysis.

Automation of the process on the Freedom EVO has enhanced precision

“We chose the Freedom EVO because

or aspirate the extraction solvent, which

we needed a platform that could offer

is important for quantitation. To ensure

both precise tip movements and near

total confidence in our results, we run

perfect handling of a variety of solvents,

a monthly calibration procedure to

including water, chloroform, methanol

verify the automated extraction and

and hexane. The system also needed to

derivatization of a lecithin standard of

be able to accurately remove the mature

known concentration, quantifying the

red blood cell layer, and so we worked

results by GC analysis.”

with the Tecan Integration Group to

to be replaced with high throughput, automated systems, and so the next step is to gain high level accreditation from an international body for the protocol we have established. This will open the door to potential new projects at Lipinutragen and attract interest from other laboratories engaged in this type of work. Our aim is to make this platform available to other laboratories

customize the platform to mimic our

“Determining the membrane lipid profile

manual pipetting procedures, setting up

is the key to distinguishing between

and refining unique software to control

a pathological problem and essential

all the automated movements. The

fatty acid deficiency syndrome, and we

result was a liquid handling system that

are raising awareness of this concept

is far more consistent and precise than

across the medical field through training

To find out more about Tecan’s

manual pipetting, which is exactly what

courses registered with the Italian Health

Freedom EVO platform, visit

we need for lipidomic analysis.”

Ministry. These courses are attended

www.tecan.com/freedomevo

around the world, helping to standardize lipidomic analysis,” Carla concluded.

by doctors, pharmacists and biologists “Precision is essential from the outset,

working in the fields of nutrition and

To learn more about

starting with the isolation of the mature

metabolic problems, allowing them to

red blood cells. Removal of the various

extend their knowledge of the subject

Lipinutragen, go to

layers above these cells is a crucial step

for accreditation and continuing

and must be performed identically each

professional development purposes.

time, which was extremely difficult to do

I believe that our lipidomic analysis

manually. Reproducible liquid dispensing

protocol will become commonplace in

and aspiration was also challenging; with

the future, with people having an annual

automation, we can precisely dispense

assessment. To keep pace with this

the required volume of solvent or water,

demand, manual processes will need

www.lipinutragen.it/en

TECAN JOURNAL 2/2018

21

DRUG DISCOVERY

Biosensors light the way for drug development The successful treatment of

The Moulder Center for Drug Discovery

biosensors, I can identify compounds

inflammatory diseases may lie

Research in Philadelphia, USA, serves as

that activate certain antioxidant

with controlling the production

a multidisciplinary research hub for

pathways, which may have therapeutic

Temple University School of Pharmacy,

applications.”

of particular proteins, driving efforts to identify translational

one of the oldest schools in Pennsylvania. The center focuses its

Oscar continued: “The types of things

repressors for drug targeting.

efforts on addiction withdrawal,

we can now achieve in biological

Scientists at the Moulder Center

Alzheimer’s disease, cancer, anti-viral

research are increasingly complex. There

for Drug Discovery Research

agents, and cardiovascular and

are many tools to investigate different

have developed luminescence-

metabolic diseases. Dr Oscar Perez-Leal,

properties of cells, and you can use one

an Assistant Professor, explained: “My

device to study multiple things

research focuses on drug development

simultaneously. For example, it is

for inflammatory diseases, in particular,

possible to carry out multiplex studies

multiplex studies and time-

fatty liver disease – which is increasingly

involving both luminescence and

course assays for identifying

prevalent due to obesity – and psoriasis,

fluorescence, the latter being used to

and characterizing novel

both of which have a high degree of

monitor cell viability. This approach

compounds.

inflammation and related oxidative

provides much greater insight into how

stress. My primary interest is in

cells respond to drugs, but depends on

identifying translational repressors of

having an instrument that also offers live

proteins that could become new drug

cell incubation, as well as

targets. In the last few years, I have

monochromators to allow selection of

created luminescence-based biosensors

the appropriate wavelengths for

– basically living cells that will only emit

excitation and emission. The Spark®

light when, for example, the target

multimode microplate reader – which

protein is being produced – and have a

was purchased in December 2017 – is

couple of related patents. Using these

well suited to meet these requirements.”

based biosensors for protein detection, supporting

22

TECAN JOURNAL 2/2018

DRUG DISCOVERY

I t’s convenient to have a multifunctional platform to handle routine screenings, as well as more infrequent specialized applications, which generates high quality data every day of the week – it’s fantastic!

Dr Perez-Leal

“In order to evaluate the capabilities of

incubation as well. I wanted to combine

For example, I can determine when a

the reader, I borrowed a system from

everything I might need in a single

compound reaches maximum activity or

Tecan for a week to test it out,

platform, allowing me to run lots of

the lowest concentration that shows a

supported by a company rep for the first

different assays, which I simply couldn’t

therapeutic effect, and it’s great to be

three days. I did a fair amount of

do previously. For example, developing

able to observe all of this in real time. It

preparation in advance, so I could spend

live cell systems involves DNA

makes such a difference being able to

the week setting up assays, using the

manipulation and molecular biology, so

leave the plate in the reader and not

system as much as possible and learning

quantifying DNA and protein

have to transfer it backwards and

how to operate it, as well as making

concentrations using the NanoQuant

forwards to an incubator. Before the

comparisons with other instruments

Plate™ is ideal. It’s convenient to have a

Spark, I simply didn’t have this

in terms of sensitivity and speed.

multifunctional platform to handle

functionality in the lab; it was impossible

I experimented with many of the Spark’s

routine screenings, as well as more

to do this with my previous set-up. I’m

parameters that week, and I really liked

infrequent specialized applications,

very satisfied with the instrument, and

the fact that I was able to use it and

which generates high quality data every

I’ve enjoyed working with the Tecan

understand how it works fairly quickly.

day of the week – it’s fantastic!”

team – they’ve been very supportive,”

As a result, when the system was

Oscar concluded.

purchased, I already had some basic

“The control module is intuitive and easy

assay protocols set up, and have been

to use, and it’s straightforward to select

able to get a lot of data in a short space

the steps you need. The software

To find out more about Tecan’s

of time.”

arranges them sequentially so I can see

Spark reader, visit

the overall workflow from top to bottom.

www.tecan.com/spark

“It’s not uncommon for labs to purchase

One of the great benefits of the Spark is

an instrument for a particular project

being able to identify compounds with

without thinking of the wider, long-term

interesting functionality – say at the

provision. They might only need a

12- or 24-hour time point – and then

platform with luminescence capabilities,

isolate them to run time-course assays

and then discover further down the line

using different concentrations, which

that they would have liked live cell

offer a wealth of information.

To learn more about the Moulder Center for Drug Discovery Research, go to moulder.temple.edu

TECAN JOURNAL 2/2018

23

BIOTECHNOLOGY

A tailored solution to meet bioprocessing challenges Automation offers many benefits for bioprocess development involving multiple microbial strains, yet few off-the-shelf platforms are capable of combining liquid handling tasks with microscale cultivation. The Microbial Bioprocess Lab at Forschungszentrum Jülich needed a customized platform to support its research, and turned to the Tecan Integration Group for a solution that has improved its microbial cultivation workflows.

Forschungszentrum Jülich in western

“The BioLector can run up to 48

is based just 30 km away. This allowed

Germany is one of the biggest research

experiments simultaneously in

in-depth discussion between all the

facilities in Europe, with around 6,000

microplates, which is far more productive

parties involved, ensuring that the TIG

employees. The government-funded

than working with a few bioreactors that

team had a thorough understanding

organization has multiple institutes on

require continual monitoring. However,

not only of MiBioLab’s needs, but

campus, tackling a diverse range of

we still faced the challenge of sampling

also of the BioLector and how this

fields from supercomputing and physics

and dosing, which often has to be done

microbioreactor system could be

to informatics and biotechnology. The

during the night or according to the

integrated into a liquid handling

Helmholtz Innovation Lab ‘Microbial

real-time process data. You don’t

platform. TIG’s biggest challenges were

Bioprocess Lab’ (MiBioLab) is part of

want to have somebody sitting in front

to develop a system that had sufficient

the Institute of Bio- and Geosciences,

of a bioreactor 24/7, so this motivated

space to accommodate the BioLector,

and works with industry to investigate

us to find an alternative solution.

and to coordinate movements between

the use of renewable resources for

Automation was clearly the answer,

the two platforms to enable precise

fast and cost-effective production

so we spoke to a number of providers,

sampling and dosing on a shaking

of sought-after compounds. Dr. Ing.

including Tecan.”

microplate. The result was a customized

Holger Morschett, Head of MiBioLab,

workstation with a novel configuration –

explained: “The focus of our research

The Tecan Integration Group (TIG)

using the workdeck of a Freedom EVO®

is the use of microbial cell factories

visited MiBioLab to see the BioLector

150 adjacent to the BioLector system,

to convert renewable resources

and talk through the laboratory’s

combined with the liquid handling

– for example, biomass-derived

requirements, and was joined by

and robotics capabilities of the larger

carbohydrates – into sustainable value-

representatives from m2p-labs, which

Freedom EVO 200.

added products. This could be anything from chemical intermediates for the production of bioplastics to amino acids, organic acids and drugs, as well as industrially- and pharmaceuticallyimportant proteins. Our main tasks are strain phenotyping and bioprocess development, and the large number of genetically-engineered microbial strains that we need to test is a constant challenge. It’s impossible to do this in standard bioreactors, so we have transferred our protocols to a smaller scale system – the BioLector® microbioreactor (m2p-labs) – to give us higher throughput and allow easy automation.”

Left to right: Marco Oldiges, Carin Jansen and Holger Morschett with MiBioLab’s TIG system

24

TECAN JOURNAL 2/2018

BIOTECHNOLOGY

TIG developed a customized platform for MiBioLab

I ntegration of the microbioreactor with the Freedom EVO has allowed us to perform experiments in a completely different way, giving us the opportunity to develop and test processes that we simply couldn’t do before.

The clear benefits of this approach led to us purchasing a second platform in January 2018, and both systems are now in use 24/7; I often have to book several weeks ahead when I need to use one of them!”

“Working with the system is straightforward. We had some training

Holger continued: “A deciding factor

do with the BioLector, and allowed us

in choosing Tecan was the company’s

to gather the type of data you might

ability to guarantee what no other

expect from a well-equipped lab-scale

provider could. The microplate is

bioreactor. For example, we can take

sealed with a composite foil to prevent

samples and measure substrate and

contamination, and is continuously

product concentrations around the clock

shaken in the BioLector to ensure that

– something we couldn’t normally do

the cells have a sufficient oxygen supply.

with microliter volumes.”

Previously, shaking had to be interrupted

on Freedom EVOware® when the first platform arrived, and had simple workflows in operation after just four weeks, and more complex ones within three months. We were able to get our second system up and running after just one week of testing and transferring scripts. We have welcomed the support from Tecan, particularly for software

for sample extraction or dosing, leading

“Harvesting cells at the right time

integration, and know where to get

to transient oxygen deficiency and

point is not particularly easy to

help if we need it. It’s all working well,”

problems with cell viability. Of all the

manage manually, but automation

Holger concluded.

suppliers we approached, only Tecan

has provided the solution, thanks to

was able to guarantee that its solution

two-way communication between

could pierce the foils and carry out the

the microbioreactor and Freedom

appropriate liquid handling tasks while

EVO system. The BioLector measures

the plate was still shaking at speeds of

parameters such as biomass, pH

up to 1,500 rpm.”

and dissolved oxygen levels in real time, then notifies the liquid handling

“Integration of the microbioreactor with

platform of the location of wells that

To find out more about the

the Freedom EVO has allowed us to

need to be sampled or harvested, and

Tecan Integration Group, visit

perform experiments in a completely

the aspiration volume required. The

www.tecan.com/tig

different way, giving us the opportunity

Freedom EVO has everything required

to develop and test processes that we

for sample processing integrated onto

simply couldn’t do before, using robotics

the workdeck – including a centrifuge,

to fully automate tasks such as media

vacuum filtration station, microplate

preparation, sampling, dosing and cell

shaker, microplate cooler and an

separation. The integrated platform

Infinite® M200 microplate reader – so

has expanded what it is possible to

samples can be processed immediately.

To learn more about MiBioLab and see the automated platform in action, go to www.fz-juelich.de/mibiolab

TECAN JOURNAL 2/2018

25

DRUG DISCOVERY

Making big discoveries accessible to all laboratories Developing therapies to treat

Acute myeloid leukemia (AML) is an

working on two main projects; one for

rare diseases is often hindered

aggressive cancer affecting fewer

AML and the other for T-cell lymphoid

by the limited availability

than 10 in 100,000 people each year

leukemia (TLL). The AML project

in Europe. The resulting scarcity of

started as a chemogenomics screen

primary patient samples for in vitro

for a modulator in primary AML, and

testing presents a challenge for drug

we are now validating the result of this

samples, it is difficult to

discovery laboratories seeking out novel

using the various models available to

understand the fundamental

compounds to treat them. A recently

us in the lab. The second project stems

biology of these conditions or

established group within the University

from my previous work in Boston, which

screen compound libraries for

of Parma’s Department of Medicine

identified a modulator of a transcription

and Surgery aims to address this issue,

factor which is frequently mutated in

using a wide range of sample types to

TLL. We are now collaborating with a

investigate various aspects of leukemia

biotech company to validate a small

a ‘chemogenomics’ approach

and bone marrow transplantation.

molecule drug that inhibits this target,

to overcome this challenge,

Physician Scientist Dr Giovanni Roti

with the goal of translating these

working on a nanoliter scale

explained: “After spending several years

findings for clinical use.”

to identify new treatments for

at the Dana-Farber Cancer Institute

of primary patient samples.

Without these precious

drug candidates. A group at the University of Parma is using

aggressive pediatric leukemias.

in Boston, USA, I returned to Italy last

On starting out, one of the lab’s first

year to set up a new unit for cancer

technology updates was to invest

research within the Hematology Unit

in a Tecan D300e Digital Dispenser,

here at the university. My work in Boston

which it now uses in several different

focused on pediatric cancers using

applications. “When we screen small

chemogenomics – systematic screening

molecules, we need to perform titrations

of small molecule libraries against

to find their IC50 values in multiple cell

individual drug targets or target families

lines or patient samples. Although

– to identify new targets and drugs for

this is possible to do manually in 96-

aggressive diseases such as leukemia

well plates, it is very time consuming

and neuroblastoma, and our work here is

to pipette 12 or 14 different dilutions

a natural progression of that.”

of up to 10 drugs for each cell type.

The D300e simplifies this process

26

TECAN JOURNAL 2/2018

The Roti Lab takes a flexible approach

considerably, as well as allowing us to

to this research, adapting the techniques

work with smaller volumes in 384-well

it uses according to the goal of the

plates without the risk of pipetting

study and the samples available.

errors. It is also really fast, taking

Giovanni continued: “Sometimes we

between one and four minutes to set

start with a tissue sample, sometimes

up an entire 384-well plate depending

we use cell lines or cancer models.

on the number of compounds you want

But the basic process is always the

to test. We can also perform synergy

same; start by investigating the cellular

studies using this technology, which

biology, then use this understanding to

enables us to investigate many dilutions

identify and validate therapeutic targets

of two or more drugs in combination.

using both biochemical and genetic

This simply wouldn’t be possible

approaches, including CRISPR/Cas9

manually, as the time required for

knockdown if required. We are currently

pipetting would compromise the cells.”

DRUG DISCOVERY

ssentially, E we’re translating our pipeline from bench to bedside in a much shorter timeframe.

Andrea Gherli (left) and Giovanni Roti are using the D300e to screen small molecule drugs for AML

“The speed of the system also offers the

“Another feature we really like about

exciting prospect of being able to work

the Tecan D300e is that it is very user

To find out more about Tecan’s

directly with patient samples to help

friendly, and only requires minimal

inform therapies for the clinic. Our goal

training to set up even complex synergy

drug discovery solutions, visit

is to create a system for chemotherapy

studies. As a small laboratory, this is

drugs similar to the antimicrobial

a fundamental consideration when

sensitivity testing commonly used in

investing in any new technology.

clinical environments to guide antibiotic

With only two postdoctoral fellows

prescriptions. With the D300e, we don’t

and two PhD candidates – plus a

need a lot of time to set up the early

rotation of students from master’s

phase of these experiments; we can go

or undergraduate degrees – I cannot

from receiving the patient samples to

afford to have equipment that is not

testing compounds in the incubator in

accessible any time and to everyone.

just a couple of hours. If we happen to

The software automatically calculates

identify a molecule that is already in

how much of each compound is

clinical trials or for another indication,

required – and can randomize plate

we can ask permission from the local

layouts to avoid edge effects – which

ethics committee to use the drug in

means that inexperienced users don’t

patients. In a couple of cases, we’ve

have to perform a lot of calculations

actually been able to see a repeat

to work out the dilutions. Students

of what we’ve observed in vitro in

with limited laboratory experience

these patients, so it’s proving to be a

can therefore use this system without

good prediction of in vivo responses.

compromising the machine or risking

Essentially, we’re translating our

errors, which is a huge advantage,”

pipeline from bench to bedside in a

Giovanni concluded.

www.tecan.com/drugdiscovery To learn more about the University of Parma’s Department of Medicine and Surgery, go to

mc.unipr.it

much shorter timeframe.”

TECAN JOURNAL 2/2018

27

CLINICAL DIAGNOSTICS

Long-term partners in success Continuous advances in medical understanding are leading to rapid growth in the demand for diagnostic testing, making automation essential for laboratories trying to address this increase in their workloads. Many providers are also centralizing and consolidating testing to reduce costs, relying on instruments with higher throughput capacities to meet the demands of large service laboratories. Spanish biotechnology company Vircell recently partnered with Tecan to create a high throughput automated system for its VirClia® assays, building on the success of the company’s existing VirClia instrument.

Vircell, based in Granada, Spain,

we have partnered with Tecan for

is very popular, providing on-demand,

specializes in the development and

the development of automated

automated testing for a wide range of

production of ready-to-use reagents

workstations ever since. Our latest

infectious diseases not covered by the

for clinical diagnostics. The company

system – the VirClia Plus, based

major diagnostics providers. This system

was founded in 1991 by two clinical

on a Freedom EVO® platform – is

is capable of processing 96 samples in

microbiologists looking to accelerate

a continuation of this relationship,

10 hours – which is sufficient to meet

hospital-based diagnostics for better

offering infectious serology testing

the clinical needs of most facilities – but

patient outcomes, and has grown into

in a high throughput format.”

very large laboratories can require two

a successful global player, with its

or three of these instruments to meet

products in use in over 80 countries across five continents. Vircell’s assay portfolio employs a wide range of technologies – ranging from traditional cell culture to molecular diagnostics – and focuses on esoteric infectious

Tecan has an excellent reputation in the IVD market, and you can always be sure of the quality of its solutions.

diseases. Joan Anglés, Marketing Director at Vircell Spain, described the

Chemiluminescent immunoassay

their throughput demands, and so we

company’s approach: “When Vircell

(CLIA) techniques provide an ultra-

looked to develop a higher throughput

was created, the aim was to produce

sensitive alternative to classical enzyme

instrument for the same monotest strips.

assays that simplified clinical laboratory

immunoassay formats, making them

Although we considered a number of

workflows. Quality and sensitivity

ideally suited to the detection of

laboratory automation suppliers to help

were key considerations, which is why

low abundance molecules, such as

us to develop this new platform, we

the company’s founders chose to

circulating biomarkers of infectious

have been working with Tecan for over

produce all the antigens needed for the

diseases. Vircell’s VirClia assays rely

20 years, and so the company was our

development of our kits in house. This is

on an indirect CLIA method for signal

logical choice of partner. Tecan has an

still true today, and ensures that we can

generation, with all the calibration and

excellent reputation in the IVD market,

guarantee the quality of our products, as

standards included in each test strip

and you can always be sure of the

well as be more flexible to the demands

to create a ‘monotest’ format. There

quality of its solutions. We first began

of the market.”

is no need to run separate calibrators

discussing the project at the AACC

or quality controls, and labs can avoid

exhibition in 2015, then worked closely

“Even in 1991, the demand for infectious

batch testing, allowing faster turnaround

with the Tecan team through to the

disease testing was growing rapidly,

of results using the company’s benchtop

instrument launch last spring.”

and so automation was considered

VirClia instrument.

essential to help laboratories manage

The result of this collaboration is a CE-

their workflow demands. Our first

The VirClia Plus system builds on

marked, fully automated system with

automated ELISA processor was

the success of the VirClia, as Joan

an on-deck capacity of up to 216 tests.

based on the Tecan Minilyser, and

explained: “Our original VirClia system

Using an eight-channel pipetting arm,

28

TECAN JOURNAL 2/2018

CLINICAL DIAGNOSTICS

the VirClia Plus can process 96 tests

has been excellent and, importantly,

in just three hours, with an integrated

this didn’t stop once the product was

To find out more about Tecan’s

Infinite® 200 PRO multimode reader

launched. Tecan’s engineers worked

Partnering Business, visit

to allow fully automated measurement

with our staff to ensure that the first

and reporting of results. It also offers

few instruments installed at customer

automatic barcode scanning, custom

sites performed exactly as expected,

assay set-up options and the ability to

which was vital to long-term success.

pipette directly from primary sample

Now that our engineers have become

tubes, providing a flexible, reliable and

more familiar with the system, we no

effective solution for diagnostic labs

longer require this kind of day-to-day

and hospitals.

support, but the company’s service

partnering.tecan.com To learn more about Vircell and the VirClia Plus system, go to www.vircell.com

team is always there if we come across “As with any complex project – and

something unexpected, which is very

especially any instrument intended for

helpful. We are also working with Tecan

the IVD market – there were a number

to add new features and capabilities to

of technical challenges that needed

the system, so it’s certainly an ongoing

to be overcome,” Joan continued.

relationship.”

“The support we have had from Tecan

The Freedom EVO-based VirClia Plus system offers walkaway processing of 96 samples in three hours

TECAN JOURNAL 2/2018

29

ENVIRONMENTAL TESTING

Detection at the molecular level Many common food additives and pharmaceuticals make their way directly into aquatic ecosystems.

While their effects on humans are well documented, the impact on the environment and marine lifeforms is largely unknown. This has become the focus of a collaborative project involving researchers at the Karlsruhe Institute of Technology, which has adopted fluorescence polarization to explore the problem at the molecular level, screening compounds for their affinity to nuclear receptors.

The Karlsruhe Institute of Technology

interactions on a nanoscale and their

of Tübingen, KIT and biotechnology

(KIT) in Germany was established in

applications for macro-level systems.

company Biametrics. The collaboration is

October 2009 following the merger

Siegfried Hohmann, a PhD student in

investigating the effects of food additives

of the Karlsruhe Research Center and

the analytical biochemistry group at

and pharmaceuticals – including their

the University of Karlsruhe, with the

IFG, said: “My research forms part of

metabolites and decomposition products

purpose of developing sustainable

Effect Net, a multidisciplinary project

– on aquatic ecosystems, as well as

solutions to tackle today’s societal,

on behalf of the Baden Württemberg

raising social awareness of the problems

industrial and environmental challenges.

Water Network, which brings together

these micropollutants can cause, in

KIT’s Institute for Functional Interfaces

around 20 researchers from across the

collaboration with consultancy firm

(IFG) studies solid matter surface

University of Heidelberg, the University

ÖKOnsult.”

Left to right: Sarah Roth, Siegfried Hohmann and Florian Durm with the Spark reader

30

TECAN JOURNAL 2/2018

ENVIRONMENTAL TESTING

Siegfried continued: “The aim of the project is to analyze the impact of these chemicals, from the molecular receptor level – which I am researching – through to the microbiome and behaviors of fish and snails. My particular focus is on four main compounds: the artificial sweetener sucralose, the anti-diabetic medicine metformin and

The platform can also read 384-well plates… which allows us to include our reference substance and three other unknowns in a single run, using 16 different concentrations in triplicate.

two antidepressant drugs, fluoxetine and citalopram. These substances are poorly digested by humans, so large

“We decided to use FP for fast screening

uses testosterone, which has a low

amounts end up in wastewater plants.

of samples, to identify compounds for

affinity for the estrogen receptor, and

Unfortunately, they are not effectively

more in-depth study by surface plasmon

zearalenone, which is an estrogenic

removed at this stage – further cleaning

resonance spectroscopy (SPR). We

metabolite that shows a strong affinity.

and filtration would be required – and

needed a platform that could offer

Results for varying testosterone and

enter the environment as a result.”

quick reads and, after looking at other

zearalenone concentrations were used

instruments on the market, we decided

to create a competitive plot, showing

“The nuclear receptor affinity of these

to purchase a Spark multimode

the relationship between concentration

different substances, their metabolites

microplate reader with enhanced Fusion

and the polarization of the emitted

and any products arising from UV

Optics. The system’s software is easy to

light. The half maximal polarization shift

decomposition can be determined

use, and my experience trying to set up

concentration (IC50) is characteristic

against reference standards with

the experiments on an older instrument

of each substance and its affinity, and

known affinity. A key technique for

meant I was already familiar with the

can be referenced against an estradiol

determining this affinity is fluorescence

way microplate readers work and which

standard. As expected, testosterone

polarization (FP). The principle of the

parameters could be optimized. The

showed no change in polarization as the

technique is fairly straightforward. A

platform can also read 384-well plates

concentration varied, while there was

sample is mixed with a receptor bound

– a deciding factor – which allows us

a clear decrease in polarization as the

to a fluorophore-labeled analog of the

to include our reference substance and

concentration of zearalenone increased.

compound of interest. If a compound

three other unknowns in a single run,

Overall, the technique has worked

present in the sample has a higher

using 16 different concentrations in

really well, and I’m looking forward to

affinity than the labeled analog, it will

triplicate. The readings can be taken in

adapting the method to support my

displace it and preferentially bind to

10 to 15 minutes – just enough to grab

research into micropollutants,”

the receptor. Polarized light is then

a bite to eat – in contrast to SPR, which

Siegfried concluded.

used to excite the fluorophore, and the

can take almost a day to give a single

difference in polarization between the

measurement, and is better suited to

exciting and emitted light is measured.

in-depth studies to determine kinetics

To find out more about Tecan’s

The key variable is the speed of rotation

and time-resolved measurements for

Spark reader, visit

of the labeled analog; when bound to

compounds identified through

www.tecan.com/spark

the receptor, rotation is slow, but after

FP screening.”

®

displacement, rotation is much faster.

To learn more about the

The difference in polarization between

“We purchased the Spark at the

excitation and emission is therefore an

beginning of the year, and initially

indicator of the level of displacement –

validated an assay involving estrogen

as more labeled analog is displaced, the

receptors, which are inexpensive

emitted light becomes less polarized.”

and easy to stabilize. The assay

Institute for Functional Interfaces, go to www.ifg.kit.edu

TECAN JOURNAL 2/2018

31

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Tecan Journal, Customer Magazine of Tecan Trading AG., ISSN 1660-5276 Design: OTM/London www.otmcreate.com Photography: Günter Bolzern/Zürich www.bolzern.tv Editor in Chief: Tecan Trading AG, Antonietta Allocca Editor: kdm/UK www.kdm-communications.com Editor: UP THERE, EVERYWHERE/Sweden upthereeverywhere.com Print: DAZ Druckerei Albisrieden AG/Zurich www.daz.ch Address: Tecan Trading AG, Marketing Communications, Seestrasse 103, CH-8708 Männedorf, Switzerland, hello@tecan.com, www.tecan.com Images that are not owned by Tecan have been reproduced with permission.

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Tecan Group Ltd. makes every effort to include accurate and up-to-date information within this publication, however, it is possible that omissions or errors might have occurred. Tecan Group Ltd. cannot, therefore, make any representations or warranties, expressed or implied, as to the accuracy or completeness of the information provided in this publication. Changes in this publication can be made at any time without notice.

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