Tecan Journal Edition 02/2017

Tecan Journal

Edition 2/2017

Life Sciences, Diagnostics and Partnering

Fishing for genetic information

Traditional medicine meets modern analytics

Perfect synergy for molecular diagnostics

Luciferase comes to the devil’s rescue

Pages 8-9

Pages 14-15

Pages 26-27

Pages 28-29

CEO WELCOME

Welcome Dear Reader, Here at Tecan, we are committed to providing our customers with best-in-class laboratory automation solutions that combine cutting-edge performance with exceptional reliability. This is what the Tecan brand is known for in the marketplace. It is also why leading diagnostics and life science instrument providers trust Tecan Partnering to help them build some of the most successful products on the market.

Our recently announced agreement with IVD specialist DiaSorin is a good example of the trust our partners place in us. Our FluentÂŽ Laboratory Automation Solution will be used in combination with the DiaSorin LiaisonÂŽ MDX PCR instrument to provide customers with a complete sample-to-result system for molecular diagnostics. The Fluent platform is already popular for drug discovery and clinical research applications (pages 20-23), and this will be the first time it features in an OEM project.

This issue of the Tecan Journal contains several more examples of how Tecan Partnering can help customers to accelerate and simplify instrument development, offering everything from complete automation of a proprietary technology (pages 26-27) to supply of liquid handling robotics for easy integration into a customer-developed solution (pages 10-11). These OEM development capabilities perfectly complement our portfolio of life sciences automation solutions, and it is thanks to this in-depth application knowledge and broad expertise that we remain the partner of choice for research laboratories and leading instrument manufacturers around the world.

Dr David Martyr CEO

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TECAN JOURNAL 2/2017

CONTENTS

Contents 2

CEO Welcome

4 - 7 The time is now – automation for the academic laboratory

8 - 9

Fishing for genetic information

10 - 11 OEM instrument design offers automated patch clamping solution

12 - 13 8-9 Fishing for genetic information

Working towards cancer-free childhoods

14 - 15 Traditional medicine meets modern analytics 16 - 17 Accelerating R&D through collaboration and automation

18 - 19

Taking the guesswork out of drug development

20 - 21 What is normal? Understanding the vaginal microbiome

22 - 23

Flexibility for speed

24 - 25 Faster processing of samples in steroid hormone testing

26 - 27 Perfect synergy for molecular diagnostics 28 - 29 Luciferase comes to the devil’s rescue 30 - 31

30 - 31 Expanding the experimental space

Expanding the experimental space

TECAN JOURNAL 2/2017

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BLOG

The time is now – automation for the academic laboratory

Pluripotent stem cell

The Blog 4

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

TECAN JOURNAL 2/2017

BLOG

‘There is no time like the present’

think about the day-to-day

perfectly applies to the topic we are

challenges you face, and

about to discuss: integrating automated

what it takes to keep

solutions into the workflows of

everything operating

academic laboratories. The oft-repeated

smoothly and efficiently,

and stale arguments made against the

producing high quality data.

need for automation in an academic

If you break down your typical

setting are dated and, in most cases, no

processes and protocols into individual

longer relevant. There are myriad

steps, it will become increasingly clear

advantages to introducing automated

where there are gaps and potential

liquid handling, detection and control

sources of error, and where you can gain

systems to any lab – whether it’s a

the most from improving accuracy,

single, dedicated instrument designed

throughput and efficiency. Whether your

to carry out a specific labor-intensive

laboratory is focused on genomics or

task, a modular system that can be

proteomics research, doing cell-based

reconfigured and expanded as needed,

assays, ELISAs or next generation

A point well taken. Lab technicians,

or a complete workstation with

sequencing, automation can help you

postdocs and graduate students are all

integrated components to automate

to meet your individual needs.

quite capable of manually pipetting

throughput, but it also improves pipetting accuracy and precision – the two pillars of data reliability.

“I don’t need robotics. I have enough staff to do everything manually.”

liquids, plating samples, feeding cell

your entire workflow. We will explore these options and discuss what your lab

“I don’t do the kind of high

cultures and performing assays.

could gain in terms of precision,

throughput experiments that can

However, the major problem with any

reliability, data quality, reproducibility

benefit from automation.”

manual activity is the risk of human

and productivity, as well as try to dispel

Automation doesn’t just process

some of the myths that many people still use to convince themselves that automation has no place in academia.

The elephant in the room – cost The first argument is that ‘automated instruments are too costly – I just can’t afford them.’ Research budgets are tighter than ever, and every expense must have proven value. Not only can an experienced laboratory automation provider understand what you do and don’t need, they can also answer your questions and concerns – helping you to explore instrument and system options that best serve your current requirements – offer the flexibility to adapt to meet future needs, and meet your budgetary constraints. With this in mind, Tecan has put together a special pricing program to empower academia, making it more affordable than ever to automate your workflow.

Why fix something that isn’t broken?

samples faster. Whether you are working with 100 samples or 1,000, accuracy and precision are essential for obtaining consistently high quality results.

Reproducibility is the cornerstone of

error. This can be frustrating and lead to unexplained results, but it can also be costly in terms of time and resources if you have to repeat experiments.

Consider too: • People need to take breaks during

scientific experimentation, and an

the day, and vacations (they also

inability to reproduce results has,

daydream and have lapses in

unfortunately, led to several recent

attention)

retractions of published papers in high profile journals. Standardization of

• People get sick

experimental methods to the extent

• People might leave for new positions

possible with automation minimizes

once they have mastered a protocol,

variability between runs, contributing to

requiring the training of new staff

more reproducible results that you can present with greater confidence.

Inaccurate pipetting can have damaging effects on results and data quality across

• People get bored of doing tedious, repetitive tasks, and they are at risk of repetitive stress injuries • The accuracy and precision of manual

the workflow, whether you’re setting up

pipetting varies more than that of a

screening protocols or performing serial

robotic liquid handling platform (and

dilutions. Poor pipetting performance

you cannot simply recalibrate your

can have major consequences for highly

technicians periodically!)

sensitive applications such as NGS library preparation – with its many tedious and error-prone steps – and can result in time-consuming and expensive

You may be convinced that your

resequencing.1 Automation of library

laboratory is running well, and cannot

preparation minimizes errors, reduces

really benefit from automation. But

hands-on time and enables higher

www.tecan.com/blog TECAN JOURNAL 2/2017

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BLOG

control, especially for extended time course experiments.3 Stem cells and induced pluripotent stem cells are especially sensitive to changes in temperature, humidity and other environmental conditions. A fully automated workflow with an integrated incubator, robotic liquid handling to minimize the risk of contamination, and a plate reader with built-in environmental controls can improve outcomes and data quality. This will not only increase your lab’s productivity, it can also save you from the cost and inconvenience of repeated experiments, wasted samples and reagents, and missed publication or presentation dates. Consider too that, if you can use walkaway automation to free up the valuable time of your graduate students and postdocs, they can be using their

Mutant stem cell

energy and expertise in more A study published in the Journal of Laboratory Automation tested the effects of a miscalibrated liquid handling system on assay performance.2 The

purposeful ways.

“Automated systems are too complicated to learn and operate.”

study involved two types of in vivo

A decade ago, that sentiment may have

biochemical assays, and demonstrated

been valid, but it is no longer the case.

that small changes in accuracy did affect

You can now choose from a selection of

assay performance. While there was no

ready-to-run, standalone instruments

appreciable change in signal-to-

and fully integrated turnkey systems,

background ratio or assay variability

many of which are available with built-in

(Z-factor), inaccurate dispensing had a

training videos, user guides and

measurable effect on compound

application protocols, as well as training,

potency results, and was ‘especially

technical support and ongoing

drastic for lower potency compounds’.

maintenance options. These intuitive,

The data showed that lower potency

automated workstations require little

compounds could easily be missed, and

more than loading of the materials,

that the assay loses resolution for high

telling the system what you want it to do

potency compounds that have similar

and pushing ‘start’, with preprogrammed

IC50 values.

software scripts to take the hassle out of setting up experiments.

Working with a living system When performing live cell assays, keeping your cells healthy and alive until the assay is completed requires

The Blog 6

“Automated systems are not flexible enough – I don’t know what my needs will be in the future.”

substantial hands-on time, careful

The open architecture platforms such as

monitoring and robust environmental

the Fluent® and Freedom EVO® combine

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

TECAN JOURNAL 2/2017

BLOG

exceptional ease of use with the ability to customize protocols as needed. No matter how you define ‘flexible’, consider the following: • Workstations can accommodate one or more liquid handling or gripper arms, with various pipetting heads to suit a broad range of applications and dispensing volumes

References 1) Sitarska, A. Precision and accuracy – two pillars of data reliability. Tecan blog. www.tecan.com/blog/precision-and-accuracy-twopillars-of-data-reliability 2) Hentz, NG, Knaide, TR. Effect of liquid-handling on assay performance. J Lab Automation, 2014, 19(2), 153-162.

3) Oberdanner, C. Great expectations for your live-cell assays? Here’s how to keep cells performing even when you’re asleep. Tecan blog. www.tecan.com/blog/great-expectations-for-yourlive-cell-assays-heres-how-to-keep-cells-performingeven-when-youre-asleep

• Platforms can be fully integrated with your choice of peripheral devices, including incubators, plate readers, shakers, mixers, etc.

• Modular instrument designs make it easy to reconfigure workstations as research needs change, adding, removing or upgrading individual components • Systems can sit on a lab bench or be freestanding to take advantage of the space below the workdeck, including robotic access to below-deck instruments, devices or storage From individual components to fully integrated solutions, there are now a variety of options to automate the workflows of academic laboratories.

Experienced laboratory automation providers understand the specific needs of the sector, and can help you to select the instruments, software and system configurations best suited to your laboratory. And, with our new pricing structure making these industry-proven automated solutions affordable for academia, the time is now. What are you waiting for?

Stem cells

www.tecan.com/blog TECAN JOURNAL 2/2017

7

MARINE BIOLOGY

Fishing for genetic information Canada’s Molecular Genetics Laboratory at the Pacific Biological Station uses DNA analysis to identify and track salmon from different hatcheries. Automated NGS has enabled the laboratory to introduce parentage-based tagging, a cost-effective alternative to the coded wire tag system.

The Pacific Biological Station in

produced salmon is incomplete, as not

the Pacific Salmon Treaty between

Nanaimo, British Columbia, is part of a

all fish are tagged. An alternative, more

Canada and the United States. To

network of nine major scientific facilities

cost-effective approach is parentage-

adhere to the terms of the treaty, it’s

operated by Fisheries and Oceans

based tagging (PBT), which involves

important to determine the origins of

Canada, the government agency

genotyping of hatchery broodstock.

both Chinook and Coho salmon in

responsible for the management of

Establishing a database of broodstock

fisheries. Historically, this has been

Pacific salmon fisheries. The oldest

genotypes from each hatchery allows

done using CWTs, but can now be

fisheries research center on the Pacific

the offspring to be non-invasively

achieved with PBT technologies. PBT

coast, it is home to the Molecular

sampled to identify the hatchery and

gives a more refined estimate of stock

Genetics Laboratory (MGL), which is

brood year according to parentage.

compositions than using CWTs, helping

responsible for determining Pacific

Once complete, all juvenile fish are

to determine the contributions of each

salmon population structure and

effectively tagged and can be traced to

hatchery to commercial and

estimating stock compositions in

a specific hatchery.

recreational fisheries. If there is concern about conservation of a particular

mixed-stock salmon fisheries.

The abundance of Chinook and Coho salmon in British Columbia has been enhanced by production from hatcheries, with a portion of the juveniles marked with coded wired tags (CWTs) prior to release. If an individual containing a CWT is recovered from a

I can foresee that [NGS] is going to become the way of the future in a lot of applications.

stock, knowing the timing and location of the stock enables the closure of specific areas at appropriate times.”

Terry continued: “Previously, stock composition was primarily determined by microsatellite analysis, and this technique is still a major workhorse in our lab, particularly for those

fishery, its age and hatchery of release can be determined. However, tagging

The MGL has been using DNA analysis

applications where we need an answer

with CWTs can be expensive, the tag

since the early 1990s, as research

within 24 hours. During the fishing

cannot be retrieved without killing the

scientist Terry Beacham explained: “The

season, we use microsatellites to

fish, and identification of hatchery-

catch of Pacific salmon is governed by

analyze several thousand Pacific salmon in just a few weeks, estimating the stock composition to help decide when a fishery should be opened or closed.

More recently, we have implemented automated NGS protocols, allowing hundreds of single nucleotide polymorphisms (SNPs) to be costeffectively analyzed. When we first considered using SNPs for genetic stock identification, the number of SNPs we would need to run to generate results equivalent to that of microsatellite analysis – and the cost and throughput of the platforms available at the time – meant it simply wasn’t viable. With the advent of

The Pacific Biological Station team 8

TECAN JOURNAL 2/2017

MARINE BIOLOGY

NGS protocols, we invested in an Ion

sequencing verified, flexible protocols

Proton™ System – enabling us to

for Ion AgriSeq library preparation,

cost-effectively analyze several hundred

developed in collaboration with Thermo

SNPs at one time – and have developed

Fisher Scientific. One of our technicians

methods to monitor Chinook and Coho,

attended a Freedom EVOware® training

the main species of salmon produced in

course, and the knowledge she gained

hatcheries in British Columbia.”

from this allows her to write many scripts for users from different groups,

To take full advantage of the

which is a real benefit. We’ve been

throughput capabilities of the Ion

running the systems for about two

Proton System, and to provide

years now, developing the workflow

additional flexibility in the lab, the MGL

and optimizing the scripts for our

invested in two Freedom EVO® 100

applications, and can now run four chips

workstations. “Typically, we receive 100

a week – representing over 1,500 fish –

adipose fin punches attached to a

which is a fairly good throughput.

Whatman filter paper. DNA is extracted

While NGS is quite a new technology

from these punches either manually

in fisheries management, I can foresee

using the Chelex method or automated

that it is going to become the way of

on a BioSprint instrument, ready for

the future in a lot of applications,”

Applied Biosystems AgriSeq™ library

concluded Terry.

Fisheries and Oceans Canada is responsible for monitoring the Pacific salmon population

preparation. Initially, DNA is amplified and normalized to a concentration of

All Tecan products mentioned are for research

40 ng/µl using a Freedom EVO

use only. Not for use in clinical diagnostics.

platform equipped with an Air LiHa™,

Punches are taken from the adipose fin

before adding the primer panel to the 96-well plate using the second workstation’s MultiChannel Arm™ 96.

Currently, we look at between 300 and 350 amplicons depending on the species, but hope to extend this to 500 amplicons or more. After digestion, a set of 384 barcodes is ligated to the amplified products, and the pooled libraries are transferred to an Ion Chef™.

This generates two sequencing-ready chips for the Ion Proton System, from which we determine the genotype of

To find out more about Tecan’s genomics solutions, visit www.tecan.com/genomics To learn more about the Fisheries and Oceans Canada Pacific

the fish.”

Biological Station, go to

“We chose Tecan because it is a well-

facilities-installations/pbs-sbp/

known brand with a good reputation and

index-eng.html

www.pac.dfo-mpo.gc.ca/science/

Samples are mounted on filter paper and sent to the laboratory

quality products. There is also the added advantage that the company offers

TECAN JOURNAL 2/2017

9

PATCH CLAMPING

OEM instrument design offers automated patch clamping solution Ion channels regulate many

Sophion Bioscience, in Ballerup,

with a cell’s membrane. A small amount

physiological processes, as

Denmark, was established in 2000, and

of suction ruptures the membrane to

among its aims was a desire to ‘take the

bring the cytoplasm and pipette

voodoo out of patch clamping’. Since

solution in direct contact, enabling the

its start-up days as a spin-out from

electrode to measure currents between

NeuroSearch, its team of dedicated

10-12 and 10-9 A passing through the ion

drugs. Patch clamping remains

electrophysiologists has expanded to a

channels.”

the gold standard assay for

company of around 50 employees

investigating ion channels, but

designing automated patch clamping

“Traditional, manual patch clamping

solutions. Rasmus Bjørn Jacobsen, head

requires a high level of skill, and it can

of the applications development group,

take a year’s training to become

explained: “Patch clamping is a direct

proficient. Even then, throughput is

measurement technique used to

resulting in ion channels

limited to five to ten assays per day,

investigate the passage of ions across

and each practitioner has their own

remaining poorly understood.

cell ion channels. The whole-cell

equipment, which raises concerns

method uses an electrode-containing

about reproducibility. Alternatives to

glass pipette to create a gigaohm seal

patch clamping involving fluorescent

well as playing a role in many diseases, making them a target for 20 percent of registered

the manual technique requires patience, and extensive training, and has a low throughput,

The Q-Patch offers the accuracy of manual patch clamping with increased throughput and reproducibility

10

TECAN JOURNAL 2/2017

PATCH CLAMPING

dyes have a much higher throughput, but suffer from low sensitivity and accuracy. With such obstacles to overcome, it made sense to design an automated solution.”

In 2004, the company launched QPatch® – a fully automated patch clamping system – into the market. Göran Mattsson, global product manager for QPatch, said: “The whole assay process is handled by the instrument, from cells to compounds to consumables. The user simply presses ‘Start’ and the experiment runs for up to five hours.

This unattended operation is an essential requirement of many pharma companies, who also demand high quality standards and reproducibility.

Three quarters of the top 20 pharma companies worldwide are now using a QPatch – or other Sophion products – for drug discovery and safety testing, and it has become the benchmark for

The Xantus can switch between multichannel and single channel pipetting

cardiac safety.”

“The QPatch brings together the

design; the power and the cables

accuracy of manual patch clamping

connect to the back end, and everything

with the high throughput of alternative

else – including the pump – resides in

methods, and can make up to 48

the robotic arm. Crucially, the Xantus

recordings in 30 minutes. Training a

can switch between multichannel and

technician on the instrument takes just

single channel pipetting, with the option

two days, and the assay process does

to cherry-pick samples, and we can

not require expert oversight. One

upgrade the system when a customer

person can prepare the cells, another

requires increased throughput.”

can start the process, and someone else can analyze the results. The built-in

“It’s been great to work with Sias’

security is also an added benefit; the

in-house engineering team and we

QPatch connects to an Oracle database,

collaborated quite closely to develop

which logs all data and is FDA 21 CFR

the QPatch. Not only did they help with

Part 11 compliant. The data cannot be

the integration of the Xantus robot,

deleted or overwritten, providing

they also designed a special titanium

reassurance to pharma companies that

oxide coating for the pipette probes to

they can trust the data recorded

help prevent compounds sticking to

anywhere in the world.”

them. Sias set the bar for customer

P artnering with Tecan allows us to deliver the complete service package, not just a high quality instrument.

service support by providing valuable Jørgen Due, head of service, continued:

training to our engineers, and this

“Patch clamping requires the utmost

standard has continued under Tecan. In

precision and accuracy, and is extremely

the future, if we are bringing another

sensitive to electrical noise given the

product to market, then I’m sure our

tiny currents we are measuring. When

strong relationship is set to continue.

designing the QPatch, we looked at

Partnering with Tecan allows us to

what OEM liquid handling solutions were

deliver the complete service package,

available on the market, and we came

not just a high quality instrument,”

across the Sias® Xantus® robotic

Jørgen concluded.

pipetting platform from Tecan. The Xantus fulfilled our specific needs, and

All Tecan products mentioned are for research

we appreciated its self-contained

use only. Not for use in clinical diagnostics.

To find out more about partnering with Tecan, visit www.tecan.com/partnering To learn more about Sophion Bioscience, visit www.sophion.com

TECAN JOURNAL 2/2017

11

DRUG DEVELOPMENT

Working towards cancer-free childhoods Developing cancer drugs for clinical trials involves not only identifying and evaluating suitable agents, but also observing how they interact with the cocktail of other drugs in a cancer treatment regime. For the Telethon Kids Cancer Centre in Perth, Western Australia, increasing throughput and reducing assay volumes are essential to save money and time in the race to beat cancer.

The Telethon Kids Cancer Centre (TKCC)

tumor cells for drug screening and

the D300e, it was a lightbulb moment.

brings together a dedicated group of

preclinical testing such as drug

This instrument has transformed the

researchers and clinicians from across

sensitivity assays, diluting drugs from

activities of our lab. Previously, working

the globe to work collaboratively

high to low concentrations to identify

together with a colleague, you could

towards developing more effective cures

effective doses. What’s important for

generate a small number of plates

to treat childhood cancer. Brain tumors

us is looking at how different drugs

working solidly for three to four hours.

are the most common form of solid

interact with each other; if we were to

With automation, our throughput went

tumor in children, affecting 200 children

put a new drug into an existing

through the roof and increased by four

in Australia each year, and many more

protocol, would the drug interfere with

or five times; we are now able to

worldwide. Within the TKCC, the Brain

the current treatment?”

generate faster and more consistent

Tumour Research Program (BTRP) –

results. Working manually, we would

headed up by Dr Nick Gottardo and

Three years ago, the team looked to

often see a lot more inter-experiment

Dr Raelene Endersby – is striving to

develop the automation in its workflow,

variability than we now do with the

improve patient survival rates and

to help carry out the thousands of

D300e, where all of the data overlays

quality of life, through basic and

necessary drug tests. Raelene continued:

beautifully. The error bars are much

preclinical research, with a key focus on

“We were doing lots of dilutions, and

smaller compared with our previous

providing the necessary evidence to

serial dilutions, which we were carrying

data, and the increased reliability means

help a therapy transition to clinical trials.

out either manually, or using a much

that we don’t need to repeat the

slower robot. When we found out about

experiment as many times.”

The TKCC is home to a diverse range of researchers from academia and industry, as well as clinical oncologists, neurosurgeons, radiologists, chemists, pharmacologists and bioinformaticians.

It has a close connection with oncologists at the Princess Margaret Hospital – the only children’s hospital in the state – and this relationship is key to defining the direction of its research, based on the lack of treatment options currently available to patients. The aim is to uncover more targeted, less DNA-damaging treatments to tackle the tumors.

Raelene explained: “My lab is interested in looking for agents that sensitize cancer cells to conventional treatments, such as chemo- and radiation therapy, and then evaluating them for safety and efficacy. We generate in vitro and in vivo models of pediatric brain cancer using surgical specimens or genetically modified mice, and then use those 12

TECAN JOURNAL 2/2017

Left to right: BTRP team members Tracy Seymour, Hetal Dholaria, Stacey Fazio, Brooke Strowger, Raelene Endersby, Mathew Ancliffe, Hilary Hii and Jacqueline Whitehouse

DRUG DEVELOPMENT

Dr Jacqueline Whitehouse, senior

“It’s very easy and quick to set up; you

scientist in the BTRP, agreed: “What

just copy and paste your data from

would take hours on our previous liquid

Excel®. It’s been simple to train people

handling robot, now takes minutes on

and there’s not much people can do to

the Tecan system. The fact that we can

break it; the leukemia group within the

use small picoliter volumes also saves

TKCC is also using our system and it’s a

money, as some of the novel anticancer

good instrument to share with people

drugs are quite expensive and hard to

knowing that it will come back in one

synthesize. Using manual pipetting or

piece. We’ve used it pretty heavily for

our previous robot, we could only work

the past two years and, since there are

with a minimum volume of about five

not many moving parts, it’s been great,”

microliters. We would be wasting a lot of

Jacqueline concluded.

W ith automation, our throughput went through the roof… we are now able to generate faster and more consistent results.

drugs just by the nature of the pipettors and their accuracy, whereas we can now

All Tecan products mentioned are for research

dispense picoliter volumes consistently.

use only. Not for use in clinical diagnostics.

We were skeptical at first, but were pleasantly surprised when we were still seeing effects of our drugs on cells using such small volumes. The other advantage is its size, it’s so small that it can easily fit inside a biosafety cabinet.

This is important for both our precious cultured cells – minimizing the risk of contamination – and for the user, protecting them from exposure to potentially hazardous or toxic novel chemotherapeutics.”

To find out more about Tecan’s liquid handling and automation solutions, visit www.tecan.com/mdx To learn more about Telethon Kids, visit www.telethonkids.org.au

TECAN JOURNAL 2/2017

13

DRUG DISCOVERY

Traditional medicine meets modern analytics Chinese medicine combines herbal remedies with acupuncture, massage, exercise and diet to provide alternative therapies for a wide range of conditions. Despite drawing on over 2,500 years of traditional knowledge, little is known about the mode of action of these herbal medicines. Researchers at Zhejiang University’s College of Pharmaceutical Sciences are looking to address this, using modern laboratory techniques to identify the numerous active pharmaceutical ingredients and synergistic effects that contribute to their efficacy.

Many modern medicines – from aspirin

The College of Pharmaceutical Sciences

or combination of components is

and codeine to the chemotherapy drug

at Zhejiang University is working to

responsible for a given biological effect.

paclitaxel (Taxol®) – are derived from

uncover the secrets of traditional Chinese

We have overcome this by isolating

natural sources. In fact, it is estimated

medicines, analyzing some of the 300+

crude extracts, then separating these into

that up to 50 percent of clinically

herbs used in these preparations.

individual components or fractions based

approved drugs since 1981 have been

Professor Yi Wang explained: “Many

on size, hydrophilicity, etc. Using this

either directly or indirectly derived from

traditional medicines used in China to

approach, we have built up a component

natural products.1 As small molecule

treat illnesses, injuries and infections have

library of over 30,000 fractions over the

pharmaceutical discovery pipelines have

over 1,000 years of history, but few have

last decade, which we can screen against

begun to falter, many companies are

been studied to identify the active

cells or specific molecular targets.”

looking towards ethnopharmacological

components. The main challenge we face

products – or traditional medicines as

is that the chemical composition of most

“For this work – which formed part of my

they are more commonly known – as a

herb extracts is extremely complex.

PhD research – we used an Infinite® F200

source of novel active pharmaceutical

Chromatographic separation of a single

multimode microplate reader to develop

ingredients (APIs). The problem with this

extract can yield hundreds or even

a large number of screening assays,

approach is that few traditional medicines

thousands of compounds, making it

including several cytotoxicity assays to

have been studied to identify their APIs.

difficult to identify which component

identify antitumor and antioxidative compounds. At that time, we chose the Infinite reader for its ease of use and automated injector module, which allowed us to quickly screen numerous components. This system has become the workhorse of the lab – it still works just as reliably today – but, on the recommendation of our local Tecan representative, we have since acquired a monochromator-based Infinite M1000 to help us perform more sensitive analyses and establish new applications.”

Professor Wang continued: “The free wavelength selection possible with the Infinite M1000 has helped us to design several fluorescent probes that can report on the up- or down-regulation of the activity of certain enzymes. For example, we have created a fluorescent probe for sirtuin 1 activity – a protein thought to play a role in intracellular regulatory processes – that could not

The group has built up a large component library to simplify screening 14

TECAN JOURNAL 2/2017

previously be monitored

DRUG DISCOVERY

T his is such a powerful tool for drug screening, allowing very accurate dispensing of extremely low volumes of compounds.

spectroscopically. We used this probe to

“The main advantage of the D300 is that

1) Newman, DJ, Cragg, GM. Natural products

test a panel of around 200 candidate

it can automatically set up synergy

as sources of new drugs over the 30 years

compounds for sirtuin 1 activation or

experiments, calculating and accurately

from 1981 to 2010. J Nat Prod, 2012, 75(3),

inhibition, and actually found 13 active

dispensing the required volume of each

311-35.

compounds – which is a very high hit rate

compound in a randomized layout to

All Tecan products mentioned are for research

compared to random screening.”

produce clean dose-response curves.

use only. Not for use in clinical diagnostics.

This is extremely difficult to perform by “Our work has taught us that many

hand, as there are accuracy and

Chinese medicines appear to rely on

reproducibility issues when performing

synergistic effects between multiple

the necessary dilutions to achieve

components, rather than a single API.

nanomolar concentrations of individual

Although there are some very potent

components. The D300 eliminates these

compounds, in many cases, the effects of

problems, especially for very sensitive

a single isolated component is weaker

cell viability assays, producing beautiful

than a comparable, internationally-

dose-dependence curves and helping us

University’s College of

registered small molecule drug. In

to select the best ratios of compounds.”

Pharmaceutical Sciences, go to

contrast, several compounds working in

To find out more about Tecan’s drug discovery solutions, visit www.tecan.com/drug_discovery To learn more about Zhejiang

www.cps.zju.edu.cn

combination can provide a much more powerful cellular response. Investigating these drug synergies can be very difficult to achieve manually, as pipetting small volumes of multiple drugs in complex microplate patterns is difficult to perform accurately and mistake-free. However, during a year working as a visiting scholar at Massachusetts General Hospital in Boston, USA, I was lucky enough to use a D300 Digital Dispenser for cancer drug screening. This is such a powerful tool for drug screening – allowing very accurate dispensing of extremely low volumes of compounds – and so, when I returned to China, it was the first piece of equipment I wanted to purchase.”

Members of the traditional medicines research group (left to right): Hao Li, Hao Liu, Yu Zhao, Shujing Zhang and Professor Yi Wang TECAN JOURNAL 2/2017

15

DRUG DEVELOPMENT

Accelerating R&D through collaboration and automation Discovering and developing

SMALTIS, established in 2014 by Dr

interactions by taking regular images

new antimicrobial drugs to

Sophie Guénard and Dr Cédric Muller, is

over the course of five hours. When we

a CRO specializing in microbiology,

started performing aggregation assays,

molecular and cellular biology. It offers

our customer gave us a specific

services to private and public research

operating procedure which was

laboratories, assisting with R&D

designed for a high content imaging

to new compounds. A range of

projects by providing customized

platform. As these systems are

tests are needed to determine

bacterial strains, evaluating the activity

expensive, we looked at alternative

the efficacy of potential

of antimicrobial compounds and

approaches, and developed a manual

analyzing microbial samples. The

set-up: we incubated the plate on a

company was born out of a shared

rotating incubator and removed it every

research background, as Cédric

hour for analysis, then returned the plate

explained: “Sophie and I studied

to the incubator. Although this worked

company in Besançon,

microbiology at the Patrick Plésiat labs,

well, it was very labor intensive.”

France, test automation

seeking to understand the antibiotic

has dramatically improved

resistance mechanism exhibited by

“We then came across a Spark®

Pseudomonas aeruginosa, the primary

multimode microplate reader in one of

cause of death in cystic fibrosis. We

the research laboratories of Jean-Minjoz

were fortunate to work with a big

Hospital that we collaborate with. We

pharma company to develop a specific

developed an aggregation assay

antibody vaccine and, thanks to the

protocol on the hospital’s instrument,

project, we got the idea of creating a

and a member of the Tecan team helped

company dedicated to helping

us to perform some validation

laboratories accelerate their

experiments to confirm that we could

microbiology R&D. We perform

transfer our technology and methods

minimum inhibitory concentration (MIC)

onto the system. We then went ahead

studies and time-kill experiments,

and purchased our own Spark reader,

characterizing how the test compounds

allowing us to perform the same

act on the target bacteria, and how the

aggregation assay far more cost-

bacteria in turn adapt to the drug,

effectively than using a high content

helping our clients to improve the

imaging platform.”

tackle antibiotic resistance requires an understanding of how bacteria respond and adapt

drugs, such as aggregation and adhesion/invasion assays.

For SMALTIS, a biotechnology

throughput and data collection, freeing up research hours to concentrate on developing new experiments.

efficacy of their compounds.”

Cédric continued: “The experiment now Aggregation and adhesion/invasion

runs without the need for supervision,

assays are among the many services

with the added benefit that the Spark

SMALTIS offers. In order to colonize and

produces growth curves of the strains

initiate an infection, bacteria adhere to

over the course of the assay. Automation

host cells via the fimbriae (hair-like

has freed up our time to develop other

structures) that interact with specific

experiments, for example, we are

host cell receptors. Aggregation assays

currently developing a method to

are used to assess the ability of a

evaluate luminescent bacterial strains,

compound to inhibit adhesion, by

created by inserting a copy of the

interacting with the fimbriae to cause

luxCDABE operon – from Photorhabdus

aggregation. Cédric said: “Our basic

luminescens – into their DNA.”

assay monitors drug-bacteria

16

TECAN JOURNAL 2/2017

DRUG DEVELOPMENT

W hat took one person three days, now takes an hour and a half.

“We also use the Spark to carry out

adhesion/invasion capabilities. Prior to

adhesion/invasion assays to determine

having the Spark, we had to take four

the capacity of bacterial strains to

to six images to cover each 96-well

attach to or invade eukaryotic cells.

plate, and analyze the colours with

The cells are infected with a bacterial

a microscope. This is now done

strain, then after a series of washes,

automatically, and what took one

the eukaryotic cells are stained with

person three days, now takes an hour

DAPI and the bacteria with specific

and a half. We are very happy with the

antibodies conjugated to an AlexaFluor®

Spark; it easily adapts to each assay

dye. We measure the blue and green

set-up, and its high throughput and

fluorescence, from the eukaryotic cells

automation has enabled us to

All Tecan products mentioned are for research

and bacteria respectively, to determine

continually improve our service for

use only. Not for use in clinical diagnostics.

a ratio reflecting the strain’s

our clients,” Cédric concluded.

To find out more about Tecan’s Spark reader, visit www.tecan.com/spark To learn more about SMALTIS, visit www.smaltis.fr

Left to right: SMALTIS team members Julie Challant, Sophie Guénard, Marjorie Robert-Nicoud, Cédric Muller (back), Elise Ponçot and Charlotte Richardot-Saura TECAN JOURNAL 2/2017

17

CELL BIOLOGY

Taking the guesswork out of drug development California-based company zPREDICTA™ has created a novel technology that reconstructs physiologically-relevant organ-specific human microenvironments that help eliminate the guesswork from drug development. Meaningful drug discovery studies involve complex experiments that are not feasible to perform manually. Automation is the answer, improving accuracy, saving time and reducing the amount of compound used.

A major problem in drug discovery and development is that a high proportion of the drugs reaching clinical trials – around 95 percent – ultimately fail due to a lack of efficacy. In part, this is because non-physiological 2D cell culture and mouse models frequently

used in preclinical drug discovery have

on the basis of a poor performance in

limited effectiveness as a representation

mouse models might actually work very

of human diseases. This can result in a

well in people. Based on the principle

poor correlation with the response

that cures for human diseases require a

observed in clinical trials; drugs

human-specific discovery and testing

demonstrating promising results in

environment, zPREDICTA, based in San

preclinical studies in physiologically-

Jose, USA, developed a novel 3D

irrelevant models may prove ineffective,

technology for reconstruction of a

while compounds that were deprioritized

physiologically-relevant, organ-specific human microenvironment, the Reconstructed Organ (r-Organ) platform, which mimics the native architecture of human tissues in an organ- and diseasespecific manner. As a result, there is a high correlation between preclinical studies and clinical response.

The r-Organ platform lies at the core of zPREDICTA’s goal to integrate different technologies and automate the various steps in drug development and establish a seamless workflow, from the initial drug screening process through to preclinical and clinical studies. Dr Julia Kirshner, founder and CEO, explained: “Our research involves three-dimensional organ reconstruction within an organspecific extracellular matrix, followed by cell culture under human disease conditions. As this process is very close to what happens in a person, when the drugs are applied to this platform they behave in the same way as they do in a patient, resulting in a high correlation with clinical response.”

Julia continued: “The r-Organ platform is quite versatile – it can handle a wide range of drug types, including small molecules, antibodies, antibody drug conjugates, and CAR-T cells – and the

Dr Niyati Jhaveri uses the D300e for drug development studies

18

TECAN JOURNAL 2/2017

technology is already starting to be used

CELL BIOLOGY

his [the D300e] T has contributed to a four- to five-fold improvement in accuracy.

The intuitive D300e software makes it easy to set up an experiment

in the pharma industry. We currently offer a fully validated Reconstructed Bone Marrow (r-Bone) for the study of

Set-up time

leukemia and multiple myeloma, and are

(including calculations and making dilutions)

establishing a lung system for the investigation of non-small cell lung

Treatment time (60 plates)

cancer. We also have other tissues at

(single/combination treatment and vehicle backfill)

different stages of development and

Time savings

validation, including brain, a lymph node for the study of lymphomas, and mammary gland tissue for breast cancer.

These physiologically-relevant tissues are ideal for drug development studies.”

“When you are performing drug testing studies, multiple steps are involved – dilution, dose-response, combination treatment – to acquire data at different doses and various time points, and the process really needs to be automated.

For instance, some larger scale projects have involved a couple of dozen patients, with 240 drugs and a seven-point dose-response curve. That’s anything from a couple of thousand wells through to 10,000 for the entire study, which is a huge undertaking manually. It is critical that there are no transfer errors, but it is very difficult to manually track every well to ensure that the contents are correct.

You also need to take variation across the plate – for example, due to evaporation – into consideration by randomization of the well contents. Manual processing is just not feasible. It is incredibly tedious and extremely difficult to do without making mistakes, and that’s why we invested in a D300e Digital Dispenser from Tecan.”

“We’ve had the D300e for about 18 months now, and find it very user friendly

Amount of compound needed Compound usage Hazardous chemical waste

Manual

D300e

3.5 hours

15 minutes

26 hours

2.5 hours

10× (3.3 FTE days) 68 μl

80 % reduction 20 ml

Waste reduction Accuracy (standard deviation)

13.6 μl

7 μl >2000×

11.4

Accuracy improvement

2.6 4-5×

Implementation of the D300e has led to a significant improvement in overall performance

and easy to operate. Automation has

“I really enjoy using the D300e and it’s

made a tremendous difference to our

a pleasure working with Tecan’s

workflows. Manual errors, such as missing

representatives, engineers and scientists.

wells or pipetting into the wrong well,

The team is very professional and we

have been eliminated, and we can set the

have established a great relationship,”

system to shake the plate after dispensing

Julia concluded.

to ensure even distribution and suspension of the drugs in the matrix,

All Tecan products mentioned are for research

preventing the creation of higher dose

use only. Not for use in clinical diagnostics.

pockets. This has contributed to a four- to five-fold improvement in accuracy. We save a great deal of time too. Manual set-up that took three and a half hours can be completed in just 15 minutes with the D300e, and the treatment time for 60 plates is reduced from 26 to two and a half hours. Other benefits include an 80 percent decrease in the amount of compound used, and more than a 2,000

To find out more about Tecan’s cell biology solutions, visit www.tecan.com/cellbiology

times reduction in waste, which will

To learn more about zPREDICTA,

become increasingly important as we

go to www.zpredicta.com

scale up our experiments.”

TECAN JOURNAL 2/2017

19

GENOMICS

What is normal?

Understanding the vaginal microbiome Microbiome research is still in

The human microbiome – the collection

interplay of microbial populations within

its infancy, with little currently

of micro-organisms living on or within

the gastrointestinal tract. However,

our bodies – plays a vital role in our

microbiota are also thought to play a

health, from assisting in the digestion of

significant role in women’s reproductive

foodstuffs within our gut to causing or

health, and so we are looking into this

preventing certain diseases or cancers.

field too. The major problem with this is

health and the genesis of

Despite this, the composition and

that, in order to understand what is

disease. Researchers at the

maintenance of the microbiome is poorly

abnormal, first we need to know what is

Karolinska Institute are using

understood. Although some progress has

‘normal’. Because so little comprehensive

been made in this area over the last

research has been done in this area, a

decade, a majority of current research

‘normal’ baseline of the micro-organism

initiatives are directed towards the gut or

population has not been established, and

skin microbiomes, with little focus on

so we have just begun a large study to

other tissues and biofluids.

better define this.”

The Centre for Translational Microbiome

“When samples arrive in our laboratory,

Research (CTMR) – a collaboration

we begin by extracting and aliquoting

between Sweden’s Karolinska Institute,

the DNA using a Freedom EVO®

Science for Life Laboratory (SciLifeLab)

workstation. The resulting extracts are

and Ferring Pharmaceuticals – and the

quantified using a Spark® reader, then

Human Microbiome Translational

one aliquot is sent to the biobank, and a

Research Program (HMTRP) aim to

second is used for sequencing. Most of

better understand the contribution of the

our studies rely on 16S rDNA sequencing

understood about the role micro-organisms play in both maintaining our day-to-day

next generation sequencing to establish a baseline of the microbiota present in healthy individuals as a starting point for the development of new therapeutic strategies for a wide range of diseases.

H uman error is one of the greatest risks to results in any lab, and so I am a big fan of using automation to reduce hands-on time as much as possible. human microbiome to human health,

to identify the micro-organisms present,

with the goal of developing novel

which starts with DNA normalization.

therapies. Its current focus is on the

This is performed by our Fluent®

microbiomes of the gut and the female

Laboratory Automation Solution, which

reproductive organs. Maike Seifert,

uses the concentration data from the

Laboratory Engineer at CTMR, explained

Spark to calculate the exact pipetting

the center’s approach: “The gut

volumes to achieve normalization. The

microbiome was the first microbial

system then performs the library

population to be investigated, and

preparation, generating sequencing-

initiatives such as the Human Microbiome

ready samples that can be run on our

Project mean we are now beginning to

MiSeq™ platform.”

understand the composition and

20

TECAN JOURNAL 2/2017

GENOMICS

Maike continued: “We chose the Fluent system for this application because our previous liquid handling platform was not able to perform eight-channel pipetting.

This meant that normalization had to be done one sample at a time, taking two and a half hours per plate and creating a major bottleneck in our workflow. In

To find out more about Tecan’s genomics solutions, visit www.tecan.com/genomics To learn more about the Centre for Translational Microbiome

comparison, our Fluent can perform the

Research, go to

same normalization in just 15 minutes,

ki.se/en/research/centre-for-

saving a huge amount of time. Another

translational-microbiome-

benefit, which we hadn’t appreciated

research-ctmr

until we began validating the system, is that the Fluent pipettes incredibly precisely; I’ve never seen an automated platform anywhere near as accurate – it’s wonderful. This will be particularly useful for whole genome sequencing, which requires extremely precise amounts of DNA from very low concentrations of starting material – as little as 0.2 µg/µl – to achieve high quality libraries.”

“Using the Fluent has certainly improved the quality of our library preparations, and our laboratory staff are happier. It has also virtually eliminated the risk of pipetting errors during normalization – a task that requires a lot of focus to perform manually. Human error is one

DNA is extracted and aliquoted using a Freedom EVO workstation

of the greatest risks to results in any lab, and so I am a big fan of using automation to reduce hands-on time as much as possible, especially for high throughput studies. We currently perform one sequencing run a week, which could potentially be done manually, but using the Fluent workstation is better for both our staff and samples, and will allow us to increase this to two runs a week as our sample numbers increase,” Maike concluded.

All Tecan products mentioned are for research use only. Not for use in clinical diagnostics.

TECAN JOURNAL 2/2017

21

BIOANALYSIS

Flexibility for speed Maintaining a flexible approach can be difficult in biopharmaceutical research; core laboratories must balance the ability to adapt to individual project requirements with the need for efficient, high throughput processing of ever-increasing sample numbers. Novo Nordisk’s Research Bioanalysis Department has adopted a semi-automated workflow, which combines the versatility to work across the company’s various research areas with accurate and reproducible testing of thousands of samples a day.

Originally established in the early 1920s

and pharmacodynamic profiles of drug

Principal Scientist, explained the

to manufacture insulin for the Nordic

candidates from across the company’s

department’s workflow: “We have a

market, Novo Nordisk has retained a

research pipeline. Samples from animal

total daily throughput of around

strong focus on the treatment of

or cell culture experiments are tested

35,000 tests, and use a combination of

diabetes. Today, this expertise in protein

with a panel of 100 to 200

fully and semi-automated set-ups to

and peptide technologies is also

immunoassays to gain a thorough

ensure rapid turnaround times, and to

being used to develop products for

understanding of their activities.

give us the flexibility to adapt to

hemophilia and other bleeding

Around 15 years ago, the laboratory

changing project priorities. We

disorders, as well as growth and

automated its ELISA procedures to

purchased a Fluent® Laboratory

hormone replacement therapies.

increase throughput, and improve the

Automation Solution last year to

accuracy and reproducibility of testing.

further increase our throughput, and it

Biopharmaceutical development and biomanufacturing rely on an in-depth

The sensitivity and convenience offered

understanding of the complex in vivo

by bead-based AlphaLISA®

actions of therapeutic products to

immunoassays have made it the

ensure their safety and efficacy. The

technology of choice within the

Research Bioanalysis Department,

department, and these

based at Novo Nordisk’s research

chemiluminescent assays now account

center in Måløv, Denmark, is responsible

for over 70 percent of the total

for investigating the pharmacokinetic

immunoassay workload. Johannes Fels,

is now used to perform various liquid transfers and sample dilutions as part of both our automated and semiautomated workflows. AlphaLISA is a very easy technology to run in a relatively high throughput format – there are only two reagent additions and two incubations – and so, although many of our protocols could be completely automated on the Fluent,

S taff like the Fluent platform – it is completely reliable, very fast, accurate and nice to work with. semi-automation frees up the platform for other users to perform fast assay set-up or dilutions during the incubation steps. This way, each plate only takes two or three minutes, occupying the liquid handling workstation for just a short time.”

“A typical semi-automated AlphaLISA Left to right: Kirsten Jensen, Anita Svendsen and Johannes Fels with the Fluent platform

22

TECAN JOURNAL 2/2017

protocol starts with the Fluent diluting

BIOANALYSIS

samples 10-, 20- or 40-fold from mother

can perform 100-fold dilutions in a single

to daughter plates. Next, the Fluent is

step, something we have not seen on

used to transfer samples from both

any other liquid handling system.”

mother and diluted daughter plates into a 384-well assay plate containing two replicates of each source plate. After this step, the assay plates are manually transferred to a benchtop dispenser, where assay buffer containing biotinlabeled antibodies and AlphaLISA acceptor beads is added. By running both the mother and daughter plates, we can more than double the experimental window for each assay, helping us to overcome large variations in sample or analyte concentration without performing repeat testing.

Plates are incubated offline for one hour, and then moved back to the benchtop dispenser for addition of streptavidin donor beads, followed by another 30 minutes incubation. Finally, the plates are transferred to a reader for

“Staff like the Fluent platform – it is

To find out more about Tecan’s Fluent, visit www.tecan.com/fluent

completely reliable, very fast, accurate and nice to work with – and everyone in

For more information on Novo

the department runs the same set of

Nordisk, go to

universal scripts, simply changing the

www.novonordisk.com

dispense volumes, dilutions, matrices and buffers between the various assay procedures. Everything we need is programmed into the user-friendly FluentControl™ interface, and I especially like the way the liquid classes are built up within the software. Our local Tecan application scientist helped me to do the initial scripting, so we were able to get the various protocols up and running as we wanted within two weeks of delivery,” Johannes concluded.

All Tecan products mentioned are for research use only. Not for use in clinical diagnostics.

chemiluminescence measurements.”

“Accurate and reproducible low volume dispensing is vital to achieve reliable assay performance using this very sensitive immunoassay technology, and we worked closely with the local Tecan team to design a site acceptance test that would demonstrate the suitability of the system for our needs. We were immediately able to achieve very low CVs of 2-3 % for 1 µl plasma transfers into dry 384-well assay plates, so we were confident that it would perform as expected. Another nice feature is the extended volume tip adaptor, allowing us to combine four channels of the Multiple Channel Arm to perform 96-well dispensing of up to 500 µl at a time. This makes it very versatile; you

Kirsten Henriksen (left) and Annette Larsen return samples to the hotel

TECAN JOURNAL 2/2017

23

CLINICAL BIOCHEMISTRY

Faster processing of samples in steroid hormone testing Mass spectrometry is now routinely used for clinical diagnostics around the world, but manual sample preparation and traditional liquid-liquid extraction techniques are very time consuming, requiring new approaches to help streamline laboratory workflows. The endocrinology service at Charing Cross Hospital is using high throughput automated SPE to accelerate mass spec sample prep, improving turnaround times and freeing up staff time to develop new assays.

This platform – supplied as part of a partnership between Tecan and Waters to streamline mass spectrometry workflows for clinical laboratories – has all the hardware, labware and software scripts needed to perform SPE using 96-well Oasis® MAX µElution plates.

Emma continued: “For most assays, patient samples and internal standards are pipetted into an initial 96-well collection plate on the workdeck of the Freedom EVO, then shaken for the mixing step. For scripts that include a protein crash step, we remove the plate and transfer it to a standalone Members of the endocrinology team (left to right): Somia Janjua, Anna Kowalka, Amal Bashir, Emma Walker, Sinead Reidy, Aileen Bulosan Collado and Anne Rogers

centrifuge. If the 96-well assay plates require conditioning prior to samples being added, the platform performs these steps while the collection plate is

Imperial College Healthcare NHS Trust is

spectrometry in 2008, when we

made up of five hospitals across north

converted from radioimmunoassay to

west London, UK, and treats in excess of

LC-MSMS for vitamin D testing. This was

1.5 million patients every year. The trust

originally to increase throughput, as well

also plays host to North West London

as to move away from the use of

Pathology, a consortium of laboratories

radioactive reagents. We initially

providing a wide range of pathology

continued to prepare vitamin D samples

services across its catchment area and

manually, using a liquid-liquid extraction

beyond. The endocrinology service,

approach, but the exponential increase

means that all we have to do is place a

based in the Clinical Biochemistry

in vitamin D test requesting we were

cover on the plate and transfer it from

Department at Charing Cross Hospital,

seeing at that time meant that it was not

the Freedom EVO to the mass

is an example of the specialty services

sustainable. This prompted our decision

spectrometer and begin the analysis.”

on offer, and provides advanced

to switch to a 96-well microplate format

diagnostic testing for a range of

and automated SPE (solid phase

“The Freedom EVO platform is intuitive

hormonal diseases.

extraction) for mass spec sample prep,

for the operator to use; its graphical

and we now use this approach for as

interface is compatible with a busy

LC-MSMS technology is at the forefront

many of our LC-MSMS assays as

clinical laboratory. The first assay to go

of the endocrinology laboratory’s

possible. In 2015, as part of a tender for

live using the Tecan system was our

workflow, as Dr Emma Walker,

a new mass spectrometry system, we

aldosterone testing, and we initially used

Consultant Clinical Scientist and Trust

acquired a Freedom EVO® workstation

a pre-programmed workflow. We often

Lead Healthcare Scientist, explained:

to help us further increase the efficiency

obtain basic ‘starter’ scripts from Waters

“We first began using mass

of our automated sample preparation.”

for new assays, then adapt these to our

24

TECAN JOURNAL 2/2017

being centrifuged. It then prompts the user to put the spun collection plate back onto the workdeck, and all the subsequent steps are carried out automatically, including sample addition, washing and elution into an LC-MSMS compatible solvent. This

CLINICAL BIOCHEMISTRY

MS sample prep workflow

Sample accessioning

Protein precipitation

Liquid-liquid extraction

Dilute and shoot Hydrolysis Positive pressure SPE or vacuum 96-well plate/ cartridge

Applications

Sample tracking

Chromatography and mass spectrometry

Extraction

Data integration LIMS

Drug screening and vitamins Therapeutic drug monitoring and drugs of abuse Steroids and catecholamine

Optional devices • Evaporator • Centrifuge • Sealer

specific requirements as we work

able to use to develop new assays and

through our own method validation in

widen our test repertoire. By

the laboratory. Several members of the

implementing an automated, integrated

laboratory team have been on training

approach, we have gone from a situation

courses to help us fully exploit the

where we didn’t have enough pairs of

potential of the system and we now also

hands in our laboratory, to one now

design and set up our own scripts. For

where we are using Tecan solutions to

example, we now offer a ‘core’ steroid

help us maximize what we can get out

panel, which includes testosterone,

of our mass spectrometry systems,”

androstenedione and

Emma concluded.

17-hydroxyprogesterone. We run this steroid panel assay two or three times a

All Tecan products mentioned are for research

week, with each full plate

use only. Not for use in clinical diagnostics.

accommodating around 80 patient samples – plus standards and controls – and it takes approximately 90 minutes to prepare for analysis.”

“The walkaway automation offered by the Freedom EVO workstation has saved a lot of staff time in the laboratory compared to when we were doing manual liquid-liquid extraction. Using this platform to help streamline the flow of samples through our service has

U sing this platform to help streamline the flow of samples through our service has liberated staff time, which we have been able to use to develop new assays and widen our test repertoire.

To find out more on Tecan’s clinical laboratory solutions, visit www.tecan.com/diagnostics For more information on North West London Pathology’s laboratory services, including diagnostic endocrinology, go to pathology.imperial.nhs.uk

liberated staff time, which we have been

TECAN JOURNAL 2/2017

25

GENOMICS

Perfect synergy for molecular diagnostics Automation has an important

Molecular diagnostics is becoming

gastrointestinal diseases – and

role to play in molecular

increasingly common in laboratories

microfluidics and optics expert

worldwide, and this has driven a demand

Genewave in 2013 to form the present

to move from tedious, time-consuming

day organization. Juha Kirveskari, R&D

and labor-intensive manual processes to

Director at Mobidiag, explained: “Clinical

automated workflows. A further

laboratories performing stool diagnostics

throughput. Mobidiag has

consideration for clinical laboratories is

can be working with dangerous

developed a CE-marked,

the potential hazards associated with

pathogens. User safety is paramount,

automated platform for

handling dangerous pathogens, where

and molecular diagnostics can minimize

user safety is a priority.

the risk associated with these analyses.

diagnostic workflows, minimizing manual interventions to help enhance

nucleic acid extraction and PCR plate set-up, enabling high volume screening and antibiotic resistance testing for gastrointestinal pathogens.

However, when performed manually, the Mobidiag is a Finnish biotech enterprise

process is laborious and time consuming.

specializing in developing and marketing

We already had a semi-automated

novel, innovative solutions for molecular

platform, and wanted to streamline the

diagnostics of infectious diseases. The

process further, fully automating our

Espoo-based company was originally

suite of real-time PCR-based assays for

founded in 2000, merging with

gastrointestinal diseases to eliminate the

Amplidiag – a stool-based clinical assay

numerous manual procedures required

development company focused on

before and after extraction.”

The Amplidiag Easy R&D team (left to right): Mikko Vainio, Anniina Raitila, Juha Kirveskari, Sanna Laakso and Jaakko Kurkela

26

TECAN JOURNAL 2/2017

GENOMICS

T he automated system enables us to perform the extraction in two hours, from start to finish, producing two ready-to-run qPCR plates.

Mobidiag worked with Tecan to develop

48 samples at a time, physically

On completion of the run, automated

the Amplidiag® Easy system for

separating them in a 96-well plate.

data analysis is performed, and the

automation of nucleic acid extraction

The automated system enables us to

results exported to LIMS. Manual

and PCR plate set-up for the Amplidiag

perform the extraction in two hours,

interventions are virtually eliminated.”

multiplex stool diagnostic tests for

from start to finish, producing two

gastrointestinal infections. “I knew from

ready-to-run qPCR plates. It is just not

“As a CE-marked instrument, the

clinical experience that the existing

practical to do this manually, as it is

configuration and coding of the system

technologies had limitations, and were not

quite a complex assay set-up. In theory,

are locked, but there’s still quite a lot of

ideal for our purposes. I’d used a Tecan

it would take a full day to perform, but it

flexibility for customization, such as

platform in an open configuration in a

would be almost impossible to maintain

tailored protocols for extraction or

clinical laboratory in the past, so I knew

the high level of concentration required

PCR set-up only. This offers additional

the technology and the software, and

to handle large numbers of samples

possibilities, for example, the

that the hardware was reliable and could

without making a mistake.”

implementation of in-house assay

withstand routine daily operation without

procedures. The system is now being

technical issues. I also knew that the local

“The first stage of the workflow is

distributed across Europe and other

service personnel are highly professional.”

transfer of the homogenized sample to

countries where CE marking is valid,

the laboratory in an automation-friendly

and we are expanding and developing

Juha continued: “We discussed our ideas

tube containing liquid medium. On

our service organization to ensure

with Tecan, and the company suggested

arrival, it is barcoded and the cap is

that laboratories have access to the

some very helpful features. The result

removed, then the entire extraction

technical advice and support they need.

is Amplidiag Easy, a versatile, fully

process is performed on the automated

It has been a very successful project,”

integrated platform operated through a

platform. The run is started via the GUI,

Juha concluded.

simple TouchTools™-based graphical user

which prompts the user to check that

interface (GUI). It includes a Te-MagS™

the set-up is correct, then the DNA

All Tecan products mentioned are for research

module for our magnetic bead-based

extraction and qPCR plate set-up are

use only. Not for use in clinical diagnostics.

assay technology, a heat block, and a

performed without the need for user

PosID™ barcode reader, which saves a lot

intervention. After manual heat sealing,

of user time by automatically scanning

the plate is moved to the CFX qPCR

all the sample tubes. The system can also

instrument (Bio-Rad) for processing,

To find out more about

accommodate all the consumables

while the sample data is transferred via

Tecan Partnering, visit

needed for our assays; reagent tubes,

either a USB stick or a LIMS. The CFX

www.tecan.com/partnering

extraction beads and PCR plates.”

system reads the plate barcode and imports all the sample data and

To learn more about Mobidiag,

“In a clinical laboratory, it is vital to avoid

protocols from the LIMS, starting the run

go to www.mobidiag.com

cross-contamination, and so we extract

automatically once the plate is in place.

TECAN JOURNAL 2/2017

27

IMMUNOLOGY

Luciferase comes to the devil’s rescue Wild Tasmanian devils are vulnerable to a facial cancer discovered in 1996 and identified as a transmissible tumor a decade later. The contagious disease originated in northeastern Tasmania and spread throughout the country, decimating the devil population and raising the real possibility of extinction. Scientists at the Menzies Institute for Medical Research, University of Tasmania, have pioneered research into the problem – drawing upon the latest developments in human immunology and bioluminescence cytotoxicity assays – in the hope of developing a vaccine to save the island’s iconic marsupial.

The Tasmanian devil facial tumor

Andy Flies, a postdoctoral research

PD-L1 is upregulated in devil facial tumor

disease is responsible for as much as a

fellow at the University of Tasmania

cells in response to interferon-gamma –

90 percent decline in the population

supported by the Morris Animal

a cytokine released by activated T cells.

since 1996, proving fatal in nearly every

Foundation and a Commonwealth grant

The PD-L1 binds to PD-1 receptors on

case of infection. In 2014, a second

co-funded by Nexvet Biopharma,

the T cells, inhibiting the antitumor

genetically distinct and independent

explained: “An organism’s immune

immune response and enabling the

transmissible tumor was discovered with

system should launch an attack on any

tumor to evade detection. In short,

similar consequences – large lumps

foreign cells, which is what makes the

appear around the mouth and head that

transmissible tumors so intriguing. The

prevent eating and lead to starvation.

cancer cells are transferred as an

The recent discovery of some disease-

allograft when the devils bite one

“The big headlines in recent human

resistant devils suggests that the species

another, and our team is trying to figure

immunology have covered checkpoint

may be rapidly evolving, but there is still

out why there is no immune response. In

no devil immune response to the tumor

a recent publication,1 we showed that

in the majority of the population.

the key immune checkpoint molecule

the production of PD-L1 acts as an invisibility cloak.”

molecule inhibitors. We’re tracking what is working in human clinical trials, and trying to reverse-engineer and adapt it for our own use. Our latest publication compared key immune checkpoint molecules in nine different species, ranging from humans to bats to Tasmanian devils.2 Despite the last common ancestor of marsupial and placental animals occurring 162 million years ago, we found a remarkable level of similarity in key regions for these critical immune molecules, suggesting that some immunotherapy or vaccine approaches for humans might also work in Tasmanian devils. The lack of devilspecific antibodies has been a real obstacle to progress in this area. Using antibodies from other species has yielded only moderate success, so we have developed around 50 monoclonal antibodies of our own against seven or eight different targets. We’re starting to gather the necessary resources to carry out more advanced immunology, beginning at a similar point to human and mouse immunology 20 to 30 years

The research team at the Menzies Institute for Medical Research with actress and ambassador Bonnie Sveen (left) 28

TECAN JOURNAL 2/2017

ago with regard to the availability of species-specific reagents.”

IMMUNOLOGY

Andy continued: “To date, the chromium-51 release assay has been the gold-standard approach to studying cellular immune responses. Target tumor cells are labeled with the isotope and

I now use the Spark for all my ELISAs, due to its speed and the option to use different wavelengths.

incubated with effector cells, and the immune response can be determined by measuring radioactivity, as the cancer cells release the isotope when they are killed. Working with radioactive chemicals is a hassle, as there are many regulations and chromium-51 has a really short half-life of only 21 days; after a single month, half of a $500-dollar isotope stock has decayed and, if you encounter a delay, you end up wasting your money.”

“There was a need and a desire to move on and find an alternative method, and that’s where the Spark® comes in. Our colleague, Nuri Guven, raised the funds to purchase the instrument, and I first became aware of the Spark’s great potential after I attended a Tecan Spark workshop organized at the Menzies Institute for Medical Research. Nuri’s previous experience with Tecan systems was very helpful in getting us up to speed on the instrument. In the new assay, I transfect the target tumor cells to express luciferase – an enzyme derived from the firefly – making them luminescent. The bioluminescence is proportional to the number of surviving tumor cells, so the cell survival rate can be measured by quantifying the luminescence. Although we have a few other readers in the department, they were not capable of carrying out this particular assay.”

“The Spark software is pretty straightforward and the results from the first two tests confirmed that it was suitable for our needs. The cytotoxicity assay is really fast; running a whole plate takes about 90 seconds, and it is much better to run eight plates in 15 minutes than waiting for a chromium counter for two hours. I now use the Spark for all my

1) Flies Andrew, S et al. PD-L1 Is Not

ELISAs, due to its speed and the option to

Constitutively Expressed on Tasmanian Devil

use different wavelengths. In the future,

Facial Tumor Cells but Is Strongly Upregulated

we plan to purchase the Humidity

in Response to IFN-γ and Can Be Expressed in

Cassette and Te-Cool®, which will enable

the Tumor Microenvironment. Frontiers in

us to incubate cells in the reader, allowing

To find out more about the Spark’s luminescence optics, visit www.tecan.com/spark

Immunology, 2016, 7, 513.

To learn more about Andy Flies’

us to run kinetic assays with automatic

2) Flies Andrew, S et al. Comparative Analysis

injection overnight. Our ultimate hope is

of Immune Checkpoint Molecules and Their

research, visit

that these new assays will help us to

Potential Role in the Transmissible Tasmanian

utas.edu.au/profiles/staff/

develop a vaccine but, whatever the

Devil Facial Tumor Disease. Frontiers in

menzies/andrew-flies

outcome, our research has implications

Immunology, 2017, 8, 581.

for our understanding of human and

All Tecan products mentioned are for research

veterinary cancer,” Andy concluded.

use only. Not for use in clinical diagnostics.

TECAN JOURNAL 2/2017

29

DRUG DEVELOPMENT

Expanding the experimental space Biomanufacturing requires careful separation of the molecule of interest from other cellular products to ensure the quality and stability of the final product. This is even more important for the manufacture of biotherapeutics, as the presence of unwanted molecules in pharmaceutical preparations can affect the efficacy and safety of biologically-derived drugs. FUJIFILM Diosynth Biotechnologies is using automated chromatography condition screening to generate more data in less time, helping to improve the performance of purification processes and accelerate biological drug manufacturing.

Biotherapeutics are now the key growth

derived medicines, and works with some

effects. This process is complex even

area for pharmaceutical manufacturers

of the most prominent biotech

for large macromolecules derived from

globally, but the complexity of

developers and pharmaceutical

mammalian cell lines, such as

manufacturing biological drugs means

companies in the industry.

monoclonal antibodies (mAbs), but is

that there are significant costs

even more difficult when using

associated with the development of

The company’s UK site near Billingham

microbial expression systems. Column

scalable production and purification

specializes in microbial expression

chromatography is the method of

processes. Rather than cultivating

platforms – mainly yeast and E. coli –

choice for most biomanufacturing

in-house expertise in production

and has developed a high throughput

applications, and so we need the ability

bioengineering and purification, many

chromatography workflow to help

to screen a wide range of

pharma and biotech companies now

select and optimize its purification

chromatography resins and parameters

partner with contract development and

processes. Jonathan Rapley, Staff

to ensure high purities and yields for

manufacturing organizations (CDMOs)

Scientist in High Throughput Process

our pharma customers. This is both

for the production of new

Development, explained: “The cGMP

labor intensive and time consuming to

biotherapeutics. FUJIFILM Diosynth

manufacture of biotherapeutics requires

perform manually and so, historically,

Biotechnologies has over 20 years of

the careful separation of the molecule

the scope of these screens has been

experience in process development and

of interest from other cellular products

limited by the development timelines

cGMP manufacturing for biologically-

and components to avoid off-target

of each project.”

Automated chromatography condition screening helps to generate more data in less time

30

TECAN JOURNAL 2/2017

DRUG DEVELOPMENT

The Freedom EVO 200 offers complete automation of resin and condition screening

T he Freedom EVO platform is a truly open laboratory automation solution; you can make it do almost anything.

Using design of experiment principles,

almost anything if you put the time into

we can fully explore the experimental

programming it.”

window, instead of having to investigate a far smaller number of combinations

“Both myself and a colleague have been

of resins and buffer conditions based

on Tecan training courses to further

on previous experience. This ensures

explore the potential of these

more complete data sets and helps

workstations, and we are now the ‘super

our project managers to make better

users’ who program new applications

informed decisions regarding production

and support other users, which is

processes, ultimately benefitting

working really well. As more and more

our customers.”

people have begun to understand the

“Automation of chromatography

capabilities and advantages of the

condition screening is an obvious way

“The increase in the number of

systems, demand is really ramping up;

of increasing the number of parameters

experiments we can perform has

we’re increasingly running them

that can be investigated with the

obviously led to a significant rise in

overnight and at weekends to meet

available time and resources, and so in

downstream analyses, as each screen

demand,” Jonathan concluded.

2015 we looked at the options available

generates six 96-well plates of samples.

for this application. The combination of

To help us deal with this, we have also

All Tecan products mentioned are for research

the Freedom EVO® liquid handling

invested in a Freedom EVO 100

use only. Not for use in clinical diagnostics.

workstation and RoboColumns®

platform. The system is configured for

(Repligen) was an ideal solution for our

ELISA testing, but the open architecture

needs, as this would allow us to run eight

of the Freedom EVO workstation means

pre-packed miniature chromatography

that, as we have become more familiar

columns in parallel, without the need for

with the software, we have also begun

any manual intervention. We invested in

using it to automate sample preparation

a Freedom EVO 200 platform, giving us

for other analyses, such as HPLC or

enough worktable space for all the

capillary electrophoresis. This broad

reagents required to screen eight

range of capabilities is a real benefit of

different resins at three different pHs per

Tecan systems. Unlike many other

run. The increase in capacity offered by

instruments, which are restricted to

For more information on FUJIFILM

this set-up means that we are now able

single applications, the Freedom EVO

Diosynth Biotechnologies, go to

to screen far more parameters for each

platform is a truly open laboratory

www.fujifilmdiosynth.com

project within the same timeframe.

automation solution; you can make it do

To learn more about Tecan’s chromatography solutions, visit link not working. Use this one instead: www.tecan.com/ proteinpurification

TECAN JOURNAL 2/2017

31

The Blog TRENDS, NEWS, STORIES AND MUCH MORE! FROM THE EXPERTS TO YOU. www.tecan.com/blog

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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.

All mentioned trademarks are protected by law. In general, the trademarks and designs referenced herein are trademarks, or registered trademarks, of Tecan Group Ltd., Mannedorf, Switzerland. A complete list may be found at www.tecan.com/trademarks.

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