Tecan Journal Edition 02/2016

Tecan Journal

Edition 2/2016

Life Sciences, Diagnostics and Partnering

Detecting tumor ‘castaways’ could revolutionize cancer diagnosis

Seeing more clearly

A more powerful approach to biomarker discovery

HMGB1 – a silent partner in cancer?

Pages 4-5

Pages 8-9

Pages 18-19

Pages 30-31

CEO WELCOME

Welcome Dear Reader, The launch of this issue coincides with the 2016 AACC Clinical Lab Expo. This annual event gives us the chance to showcase the breadth and depth of our solutions – from standalone devices for end users to the development and support of OEM solutions through Tecan Partnering. It also provides an opportunity to demonstrate one of our key strengths at Tecan – the capacity to support customers as they move from research to clinical applications.

Our unrivalled ability to bridge the gap between these markets stems from our close ties with the life sciences research communities. Tecan solutions are empowering our customers’ research every day, and this ensures that we gain valuable experience at the forefront of emerging technologies – from NGS and mass spectrometry to liquid biopsies and single molecule detection. Combining this with our extensive workflow automation know-how, gained through many years of working in partnership with hundreds of customers worldwide, has given us an in-depth understanding of different workflows.

This experience helps us to turn our customers’ cutting-edge laboratory developed tests into bespoke automated workflows. Through Tecan Partnering, we can guide customers – from emerging companies to leading diagnostics providers – through the instrument development process, using our broad portfolio of high quality, IVD-ready components, platforms and systems to ensure a fast time to market.

Our dedicated team can guide you through system development – from early concepts and prototyping to serial production – creating fully compliant industrial and diagnostic solutions ready for global roll out. And it doesn’t stop there; we can also provide ongoing technical support, servicing, lifecycle management and staff training. This unique combination of automation expertise, commercial understanding and worldwide support is why we are the partner of choice.

Dr David Martyr CEO

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TECAN JOURNAL 2/2016

CONTENTS

Contents 2

CEO Welcome

4-5

Detecting tumor ‘castaways’ could revolutionize cancer diagnosis

12 - 13 In safe hands

6-7

Epigenetics – adding a new layer to health

8-9

Seeing more clearly

10 - 11

Igniting research on the open seas

12 - 13

In safe hands

14 - 15

Futureproofing biobanking procedures

16 - 17

Building functionality into the antibody supply chain

18 - 19

A more powerful approach to biomarker discovery

20 - 21

Consistency for a changing market

22 - 23

A modern elixir of life

24 - 25

Miniaturizing separation science

26 - 27

Automation – an extension of the analyst

28 - 29

A 3D revolution

30 - 31

HMGB1 – a silent partner in cancer?

16 - 17 Building functionality into the antibody supply chain

Introducing The Blog www.tecan.com/blog

TECAN JOURNAL 2/2016

3

CANCER DIAGNOSIS

Detecting tumor ‘castaways’ could revolutionize cancer diagnosis We may well be on the threshold of a new hope for oncology. An exciting form of analysis is poised to have a profound effect on how we manage cancer patients – in diagnostics, prognostics and drug monitoring. Automation will be key to its success.

When circulating tumor DNA (ctDNA)

miss, leading to over- or under-treatment.

is sloughed off from tumors, it can

For example, in early stage breast cancer,

be detected in liquid biopsies of just

many patients receive chemotherapy,

a few milliliters of blood. This could

even though large-scale clinical trials have

revolutionize oncology by diagnosing

shown that only 10-15 % of patients really

cancers earlier and more efficiently.

benefit from it. ctDNA tests can help to

Dr Theresa Zhang is Vice President of

identify those patients that would benefit

Research Services at Personal Genome

from chemotherapy. They can be used to

Dr Theresa Zhang, Vice

Diagnostics, and is enthusiastic about

determine whether the patient has residual

President of Research

the potential of this new form of testing.

tumor cells that are shedding ctDNA

Services, Personal

“Measuring ctDNA will help us with earlier

into the blood, which will point to a high

Genome Diagnostics

detection in the future,” she said. “It will

chance of recurrence.

be invaluable, not just in studying minimal residual disease, but also in reducing the

2. An early warning system in drug

levels of over-treatment.”

monitoring

In the lab, automation will play an essential role in these new tests. It will help to achieve cost-effective throughput, and also provide the control over contamination that is critical to find the needle in the haystack; a few mutant DNA copies shed from a tumor into the blood, surrounded by thousands, or even tens of thousands, of normal copies. ctDNA tests could change oncology in many ways: 1. Deciding suitability for chemotherapy

the body, often detects how a patient is responding to treatment earlier than conventional approaches, such as imaging.

Increased levels of ctDNA can be detected months before imaging can detect that the tumor has returned. “It’s exciting for us to detect a molecular relapse so early,” commented Dr Zhang. “This means that we know when the therapy has stopped working long before the imaging assay indicates there is a problem.”

3. Non-invasive testing for real-time information

of risk assessment can be a bit hit and

TECAN JOURNAL 2/2016

of ctDNA, released from any lesion in

surgery, then the tumor tissue is analyzed and the need for chemotherapy. That kind

4

treatment is another hot area. The analysis

For many cancers, diagnosis is followed by to help decide on the risk of recurrence

Introducing The Blog

Using ctDNA to monitor response to drug

The tumor evolves during treatment or as the disease progresses. This means that the archival tissue is not informative

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

CANCER DIAGNOSIS

enough to guide treatment decisions

in plasma, then it means you have cancer.

later on. Being able to do a test

We now know that some white blood

non-invasively, and get real-time

cells may mutate clonally and shed

information, is therefore valuable.

mutated DNA into blood as we age,

ctDNA analysis offers an excellent

but these can be easily identified.”

alternative to serial biopsies, especially for late-stage cancer patients, as their tumors tend to shed more ctDNA into

So how is it done?

Automation is key Dr Zhang believes that laboratory automation is essential when it comes to analyzing ctDNA: “Automation is key whenever a high complexity assay – such as next generation sequencing – is involved, because the techniques are too

Early last year, Personal Genome

complicated to be scaled up in a reliable

Diagnostics launched PlasmaSelect-R™.

and controlled manner. What’s more,

This automated diagnostics procedure

automation is critical for highly sensitive

analyzes ctDNA and interrogates

ctDNA tests. We need to detect one

sequence mutations, copy number

mutant molecule out of a background

Analyzing ctDNA also gives an insight

changes and translocations in a panel of

of 1,000 or, in the future, 10,000 wild

into possible mechanisms of drug

63 clinically actionable genes.

type molecules. This means that

the blood, making it easier to detect.

4. Discovering mechanisms of drug resistance

contamination has the potential to be a

resistance by the tumor. By comparing baseline, pretreatment ctDNA results

Detection is based on next generation

real problem. While automation certainly

with blood samples taken throughout

sequencing, and the company is aiming

provides cost effectiveness, it’s also the

the treatment, it is possible to determine

at CLIA approval in the near future. Many

key to controlling contamination.”

if tumors have acquired new mutations

pharmaceutical companies have already

– and if these new mutations have contributed to resistance.

Issues to be aware of

used PlasmaSelect-R for retrospective analysis of their clinical trial samples. It helps them to understand the genetic landscape of the patients enrolled in their clinical trials, which is often

Very few procedures are perfect, and

uncharacterized, since patients have

there may be some disadvantages

gone through many lines of treatment.

As a non-invasive way to detect and monitor tumors, ctDNA testing is showing great promise. But it could also have benefits beyond these applications, providing patients with greater peace of mind and less need for chemotherapy.

with ctDNA analysis. However, they are outweighed by the advantages.

Dr Zhang commented: “With the current detection approach, not all the tumors shed detectable levels of ctDNA. And if the detection method is not sensitive enough, you may miss patients. But the biological background is very low for ctDNA – it only comes from cancers.

The dogma is that if you find ctDNA

Previously, the pharmaceutical companies had to rely on archival tumor blocks taken years before the trial, which meant that they missed the real-time status of the tumor. Wherever possible, they are now using liquid biopsies to assess the molecular landscape of the cancer they are treating.

www.tecan.com/blog TECAN JOURNAL 2/2016

5

EPIGENETICS

Epigenetics – adding a new layer to health Do you feel overwhelmed with advice on what to eat and drink, or how to exercise to improve and protect your health? Pregnant women in particular are advised not to expose their developing child to alcohol, pollutants and stress.

This is sound advice, since exposure at the fetal stage can have implications for generations to come. The science behind this is called epigenetics.

Epigenetic studies are delivering a

Sweden. Harvests were unpredictable

jumped to an amazing 32 years! The

better understanding of how to tackle

for many years after, with periods of

grandfathers’ single season of gluttony

some key diseases. This has not only

crop failure and famine often followed

somehow affected their grandsons.

changed how we view the impact of

by massive harvests, and the good

our lifestyle on our own bodies but,

people of Överkalix gorged themselves

Later work showed that the paternal

remarkably, even on generations to

during these periods of plenty.

granddaughters of women who experienced gluttony or famine when

come. Epigenetics changes how genes switch on or off, without changing

Fast forward to 1984, and Lars Olov

they were in the womb or before

the DNA sequence. Nevertheless,

Bygren at Sweden’s renowned

puberty also had dramatically differing

epigenetic changes are heritable. They

Karolinska Institute was studying the

life expectancies. This was another

not only regulate gene expression in our

health of the Överkalix population.

example of what Bygren called ‘early

own cells, and predispose us to certain

Looking at births in 1905, Bygren

influences that give late replies’. His

diseases, but amazingly, these

discovered that grandsons of Överkalix

discovery clearly showed that these

modifications have now been linked

boys who had experienced good

experiences led to heritable traits, but

to disorders affecting subsequent

harvests – going from normal eating to

remained unpublished, and it was only

generations. Let’s take a look at

gorging themselves during a single

when the concept of ‘transgenerational

how this mechanism of heritability

season just before puberty, as sperm

responses’ was widely accepted almost

was discovered.

were maturing – had an average six year

two decades later that his results

reduction in life expectancy compared

received the recognition they deserved.

Feast and famine In 1800, there was a total crop failure in Överkalix, northern

with grandsons of Överkalix boys who had experienced famine during the same stage of their development. This reduced life expectancy was often attributed to diabetes and, once the data was adjusted for socioeconomic variations, the difference in longevity

What is epigenetics?

Epigenetics is the study of changes in gene expression that occur without changes in the DNA sequence. Examples include DNA methylation and histone modification, and different cells in the body have specific epigenetic profiles that modify gene expression for the cell’s function. These epigenetic changes can persist throughout a cell’s life, and even be passed on.

Gametogenesis – the creation of sperm and eggs – is of particular interest in epigenetics research.

During development, the fertilized egg forms a blastocyst containing pluripotent stem cells. These cells have the potential to form any cell type – their genetic slate has

Introducing The Blog 6

TECAN JOURNAL 2/2016

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

EPIGENETICS

been wiped clean – but epigenetic

that immune cells called regulatory

Some even

changes from either parent could

T-cells displayed asthma-inducing

contend that

potentially be passed on. Later, female

changes in expression of the Foxp3

epigenetics finally

fetuses form ovaries and egg cells while

gene, due to epigenetic tags on Foxp3

reveals the bankruptcy

still in the womb. Astoundingly, this

which could last a lifetime, and even be

of the overly-simplified

means that at one point in pregnancy,

passed on.

concept of ‘nature versus nurture’. Continued research

three generations – the pregnant ‘grandmother’, her unborn daughter

New insights, novel therapeutic

will expand our understanding,

whose ovaries are under development,

strategies

and reveal more about the role of

These examples emphasize the impact our lifestyles have, not only on ourselves, but also on our descendants. Even though epigenetic changes may eventually wear off, smoking, poor

epigenetics in normal cell function and disease. As our knowledge of epigenetics deepens, the doors will open to a whole new dimension of the mechanisms underpinning human health.

eating and toxin exposure can be carried forward in gametes.

Further reading

Understanding epigenetic gene

and the forming eggs which will then be ready to conceive children later in life – are in essence simultaneously exposed to the same environmental stresses, such as famine or gluttony.

In contrast, sperm first form during puberty in boys, and so the environment cannot start imprinting epigenetic markers until this time.

regulation opens up new possibilities

Air Pollution Causes Epigenetic

for the development of novel drugs.

Changes That May Trigger

The first FDA-approved epigenetic drug

Asthma, Discover magazine

– azacitidine – was released in 2004 for

http://discovermagazine.com/

the treatment of patients with a group of rare and deadly blood malignancies called myelodysplastic syndromes.

Azacitidine is thought to inhibit the methylation enzymes that put epigenetic tags on genes in blood precursor cells. Since 2004, the FDA

2015/sept/15-something-in-air Why Your DNA Isn’t Your Destiny, Time magazine http://content. time.com/time/magazine/ article/0,9171,1952313,00.html

has approved three other epigenetic Another example of disease-related

drugs that might work by stimulating

Dawson, MA; Kouzarides, T. Cancer

epigenetic modification was outlined

tumor suppressor genes that are

Epigenetics: From Mechanism to

by Dr Kari Nadeau, now at Stanford

epigenetically silenced when the tumor

University. She discovered that children

develops. The recent emphasis on

Therapy. Cell, 2012, 150(1), 12-27.

exposed to excessive air pollution were

epigenetics also extends beyond the

more likely to develop asthma due to

pharmaceutical industry. Nestlé has

epigenetic changes affecting genes

committed to a six year research

which prevent the immune system from

program to study how epigenetics is

overreacting to allergens, such as

involved in maternal and early life

airborne pollutants. Dr Nadeau showed

nutrition and health.

www.tecan.com/blog TECAN JOURNAL 2/2016

7

DRUG DISCOVERY

Seeing more clearly Degenerative eye diseases affect millions of people around the world, causing progressive loss of vision and, in some cases, complete blindness.

Researchers in Australia are using stem cell technologies to investigate ways of halting, or even reversing, the effects of common eye diseases.

The Centre for Eye Research Australia (CERA) was established in 1996 to investigate both the causes of and possible treatments for eye diseases, combining population health and clinical epidemiology with advanced laboratory studies. Based at the Royal Victorian Eye and Ear Hospital in Melbourne, CERA’s Neuroregeneration Research and Clinical Genetics Units are investigating potential treatments for blinding diseases – such as glaucoma, age-related macular degeneration (AMD) and inherited retinal dystrophies – using induced pluripotent stem cells (iPSCs) to generate cellular models of disease.

Associate Professor Alice Pébay, Head of the Neuroregeneration Research Unit, explained: “Obtaining ocular tissue samples from living patients has historically been a barrier to developing new treatments for blinding diseases.

The advent of techniques allowing the generation of iPSCs from adult tissue has been a major breakthrough, providing powerful new tools for disease modeling.

Using these methods, we can now derive iPSCs directly from patient biopsies, then differentiate them into specific retinal cell types to create in vitro models of the ocular condition of interest. This enables us to study the complex genetics and molecular mechanisms of diseases such as glaucoma and AMD and, equally importantly, undertake large-scale drug screening programs to find potential cures.”

The group uses the protocols developed by Yamanaka and colleagues to reprogram somatic cells taken from skin biopsies of individual patients, generating iPSC lines which form the basis of their research. “Once we had successfully created patient-derived

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TECAN JOURNAL 2/2016

DRUG DISCOVERY

The large-scale disease modeling team at CERA

stem cell lines, we needed to establish

requirements, and so we approached

our research; retinal pigment epithelium

robust protocols for the maintenance

the Tecan Integration Group (TIG)

cells – which are dysfunctional in AMD

of our cell cultures,” Alice added.

through our local Tecan offices to create

and inherited retinal dystrophies –

“This is a fairly labor-intensive and

a bespoke Freedom EVO® system

and retinal ganglion cells, which are

time-consuming process and, because

offering sterile working conditions

affected in glaucoma and other optic

our intention was to undertake

and an integrated 84-plate LiCONiC

neuropathies. This work is also being

large-scale studies – looking for

incubator. As with any new venture, it

performed on the Freedom EVO, with

potentially very small drug effects that

took us some time to develop a culture

the aim of generating and maintaining

might halt the progression of these

maintenance protocol robust enough for

the large quantities of fully differentiated cells we require for our disease modeling

We’ve made more iPSC lines in the last few months than we had over the preceding couple of years.

and drug screening programs.”

“Our ultimate goal is to find solutions for major blinding eye diseases that affect Australians, and to pioneer vision regeneration programs to give hope to people who have lost their sight. Very

pathologies – it was not practical to

use with our iPSC lines, but it is now

few labs globally are equipped to

continue performing this manually.

working well, allowing us to generate

perform this type of work, and we hope

Automation was clearly the way forward,

large quantities of cells for our research.

that this approach will lead to new

and would also help to minimize the

In fact, we’ve made more iPSC lines in

targets and novel therapies for the

variability between cell samples, giving

the last few months than we have over

prevention and treatment of common

us greater confidence that our protocols

the preceding couple of years.”

and devastating diseases. It’s very

would be reliable enough to detect even small changes in cellular behavior.”

exciting,” Alice concluded.

The automated platform and the associated offline equipment were

To find out more about Tecan’s

Associate Professor Alex Hewitt, Head

acquired thanks to generous donations

of Clinical Genetics, continued the story:

from the Joan and Peter Clemenger

“Our main aim for automation was to

Foundation and the Phillip Neal Bequest.

free up staff time and standardize the

Alice continued: “Our next task is to

cultures we produced as much as

create protocols to efficiently passage

To learn more about CERA, go to

possible. There was no off-the-shelf

and differentiate the iPSCs into the

www.cera.org.au

solution on the market that met all our

patient-specific cell types of interest to

cell biology solutions, visit www.tecan.com/cellbiology

TECAN JOURNAL 2/2016

9

MARINE BIOLOGY

Igniting research on the open seas Fluorescence measurements are helping a microbial ecologist to investigate the undersea environment, studying microbial enzyme hydrolysis in subseafloor rocks, and how microbes respond to oil spills.

world gather samples for their individual projects, performing some initial analyses on-board ship, followed by further studies back on dry land. Assistant Professor Jason Sylvan from the Department of Oceanography at Texas A&M University took part in a recent IODP drilling expedition, the first of a series of cruises to gather samples from the boundary between the ocean crust and the mantle below. Samples from these subseafloor rocks were then analyzed on board, using fluorescence-based assays to study the Subseafloor rocks play host to a range of novel microbial communities

rates of phosphatase activities in the microbial communities

The International Ocean Discovery

encountered. Jason

Program (IODP) operates scientific

explained: “People

cruises that give researchers the

have been trying

opportunity to explore subseafloor

to investigate the

sediments and rocks, and the subseafloor

boundary between

environment. Scientists from around the

the ocean crust

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TECAN JOURNAL 2/2016

MARINE BIOLOGY

and the mantle for a long time. This

phosphate, sealed, and then maintained

reader. I particularly like the temperature

expedition visited an area of the Indian

at an in situ temperature of about 10 °C.

control feature, which enables continuous

Ocean south east of Madagascar, where

The fluorescent tag remains inactive

incubation studies, taking multiple

this boundary is shallower than in most

until the phosphate is cleaved, and so

measurements over several hours. Its

other places – around 5 kilometers

the change in fluorescence is analogous

small footprint was another attraction,

below the seafloor – making it easier

with the rate of enzyme-mediated

simplifying transport between the lab

to obtain samples.”

hydrolysis. By sub-sampling and taking

and the ship, and occupying minimal

fluorescence measurements over a

space on board. The low volumes

Jason continued: “Samples are obtained

period of several weeks, I obtain an

required are also an advantage, allowing

by drilling into the rock beneath the

indication of the microbial activity. It is

measurements to be taken on even small

ocean, collecting sequential 10 meter

a very straightforward way to determine

quantities of sample. On previous cruises,

sections of rock which are then divided

the general metabolic rate for a

I used a fluorometer with a single cuvette,

into 1.5 meter sections for analysis.

microbial community that you know little

and measurements took so much longer

As a microbiologist, I have to ensure

about and, as this type of measurement

to complete. I run anything from one

that my samples are not exposed to

is quite common in microbial ecology,

plate every few days to large numbers

contamination from the air or manual

the results are comparable with those

of plates in a single day and, using a

handling, and so immediately place

of other studies.”

microplate reader – rather than taking measurements one at a time in a cuvette

one sample per core in a bag to keep it clean, rinsing it well with distilled water

“The same fluorescence method is used

– can be the difference between being

before analysis to wash off anything that

in my lab to examine how the microbial

able to do an experiment or not. With the

might be on the outside. Working in a

community responds to an oil spill,

Spark, I can set up the experiment and

filter-equipped enclosure that removes

looking at exposure to oil, and to oil plus

walk away, running a suite of samples

particulates in the air, we break open the

dispersant chemicals. In any clean-up

over the course of an hour compared to

rock and take samples from its center,

operation, the aim is to use detergents to

an entire day. And because I’m saving

ensuring that we are only examining

break the oil down into smaller droplets,

time, I can take replicate measurements,

in situ communities.”

increasing the surface area to allow

which helps to generate even better

faster microbial degradation. While the

results,” Jason concluded.

“I establish the biomass by cell counting

theory is good, it may be that some of

under a microscope, and also store

the species that you want to degrade the

samples at -80 °C for identification of

oil don’t like the detergent. Equally, other

the microbes by DNA analysis back

species may be very happy because they

in the lab. A big part of the study is

are actually eating the detergent and not

the investigation of microbial enzyme

the oil. This kind of research helps us to

hydrolysis. This involves fluorescence

understand how best to respond to any

measurements in a Spark® 10M multimode

future oil spillages.”

To find out more about Tecan’s Spark 10M reader, visit www.tecan.com/spark10m To learn more about the Department of Oceanography at Texas A&M University, visit

reader. The live sample is added to sterile synthetic sea water media along with a

“I chose Spark for these projects based

fluorescent substrate, methylumbelliferyl

on my past success with an older Tecan

ocean.tamu.edu

…because I’m saving time, I can take replicate measurements, which helps to generate even better results.

IODP drillship JOIDES Resolution (Credit: William Crawford and IODP) TECAN JOURNAL 2/2016

11

PARTNERING

In safe hands Ongoing technical support and training can be a major concern for laboratory instrument providers looking to outsource development of new platforms. Can the OEM supplier provide sufficient training for engineers and applications specialists? Will ongoing support be available for the lifetime of the instrument? And what happens if in-house engineers can’t ďŹ x a problem?

Training in this kind of low pressure environment is invaluable as a service engineer.

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TECAN JOURNAL 2/2016

PARTNERING

Effective routine maintenance and

the technical training necessary to

together and aligned correctly, and

responsive technical support are vital

install, service and repair it.”

that the differing maintenance and

for clinical laboratories, ensuring correct

calibration requirements for each system

and efficient operation of equipment

Mohammed Mattar, Customer Service

are adhered to. It really has been very

and allowing the laboratory to provide

Supervisor at Siemens Egypt, added:

comprehensive; our trainer – Roger

timely turnaround of results. As a

“Most of our training is provided by

– has been very open, answering all

result, all major healthcare equipment

other engineering staff from within the

our questions in a logical way, and has

providers have dedicated service

global Siemens service organization, but

provided the level of insight we need to

organizations that are responsible

Tecan runs a dedicated training program

effectively support this system.”

for both the upkeep of equipment at

for the Quadriga BeFree at its global

customer sites and the provision of

headquarters in Männedorf, Switzerland.

Mohammed Mattar continued: “Training

expert technical and application support

This ensures we have the appropriate

in this kind of low pressure environment is

when required. Although most of these

knowledge and technical understanding

invaluable as a service engineer, giving you

organizations will provide in-house

of the platform, and gives us a chance

the opportunity to get hands-on and really

training for their application specialists

to draw on the company’s many years

explore an instrument with the assistance

and service engineers, this may not

of experience for troubleshooting any

of a knowledgeable trainer. This helps

always be possible when instruments

issues we may encounter.”

you to develop a picture in your head

are designed and manufactured by a third-party OEM provider.

of how the machine works – and what Shawqi Mohammed Ibrahim, Field Service

issues there could be – before you get to

Engineer & Healthcare Informatics

a customer site, where the customer is

Siemens Healthineers has a long-standing

Specialist from Siemens Saudi Arabia,

obviously keen to get the instrument back

relationship with Tecan’s Partnering

took up the story: “The training is divided

online as quickly as possible.”

Business, working with the company to

into a theoretical element and hands-on

provide advanced liquid handling and

work with the instrument. The theoretical

logistics solutions to complement its

section gives an overview of the system

To find out more about Tecan’s

advanced diagnostic testing product

– looking at the important parts, what

OEM solutions, visit

portfolio. The Quadriga BeFree®

they do and how they work – then the

System is an example of how this

practical section covers how to install

partnering.tecan.com

approach benefits customers, offering

and set up the machine, as well as how to

seamless integration of a sample

troubleshoot, strip and rebuild the various

processing module with up to three

modules. As this platform is connected

BEP® III Systems to provide an effective

with our BEP III Systems, it is also very

high throughput solution for blood

important that everything is connected

To learn more about Siemens Healthineers, go to www.healthcare.siemens.com

donor screening. Frontline support for this platform is provided through Siemens’ global service network, with expert technical back-up from Tecan’s engineering and application Expertline team if an immediate solution cannot be found.

Based on Tecan’s advanced liquid handling know-how, the Quadriga BeFree includes a number of technologies not found on other instruments in the Siemens portfolio, requiring the company’s engineers to undertake specific training to ensure effective product support. Mohammed Fatouh, IB & Service Process Manager at Siemens Egypt, explained: “Although the Quadriga BeFree has been on the market for a number of years, this platform has not been available in Egypt until now. Our sales team was keen to offer this instrument to new and existing customers – ensuring we maintain our leading position in the market by providing complete automated solutions

Left to right: Mohammed Mattar, Shawqi Mohammed Ibrahim and Mohammed Fatouh at Tecan’s

– and so it was vital that we undertook

Männedorf training center

TECAN JOURNAL 2/2016

13

BIOBANKING

Futureproofing biobanking procedures The Heidelberg CardioBiobank –

range of studies aimed at improving the

then kept in cryostorage until required

part of Heidelberg University

diagnosis and treatment of numerous

for clinical or molecular studies, either

conditions. As our understanding

in house or by external partners.

in many fields – such as cardiology

We are also a core biobank for many

and oncology – has increased, many

large-scale projects and international

Hospital – collects thousands of patient samples every year for cryostorage and use in national and European medical research projects. Since moving from the

institutions have also established disease-specific biorepositories to complement the general population

main hospital campus into a new,

biobanks. One such facility is the

purpose-built facility, the biobank

Heidelberg CardioBiobank at Heidelberg

has established a state-of-the-art

University Hospital, Germany. Part of

automated workflow to improve the throughput, quality and consistency of its sample processing, helping to drive forward large-scale cardiology studies across Europe.

one of the largest internal medicine

consortia, including the European FP7 ‘BestAgeing’ consortium, the European Horizon 2020 ‘NISCI’ collaboration, and the German Centre for Cardiovascular Research’s TORCH (translational registry for cardiomyopathies) registry. In addition, we collaborate with biobanks worldwide on issues

departments in Europe, it is perfectly

such as collection techniques, sample

positioned to recruit participants for

and quality management, and the

a wide range of clinical studies, such

development of SOPs and analytical

as the investigation of novel disease

processes.”

biomarkers. Dr Tanja Weis, Head of the Heidelberg CardioBiobank, explained:

The increasing focus of genomics and

“Our main focus is the collection and

personalized medicine in many medical

Biobanks have played an increasingly

preparation of high quality biological

specialties means that the Heidelberg

prominent role in medical research over

samples – predominantly from patients

the last two decades, supporting a wide

with heart muscle diseases – which are

CardioBiobank is now responsible for the preparation and storage of over 100,000 patient sample aliquots annually. This increasing workload has meant that many of the routine sample processing activities – which were previously performed manually – have now been automated, significantly increasing the speed of its protocols, as well as enhancing the yield and quality of DNA extracted. Tanja continued: “The biobank was established about eight years ago with just a few -80 °C freezers and liquid nitrogen tanks, which rapidly grew to around 70 freezer units! However, these occupied a great deal of space and took a lot of effort to maintain. Automation became essential and so, when the biobank moved into a purpose-built laboratory on the university’s life sciences campus two years ago, we installed a fully automated LiCONiC STC cryostore with the capacity for 1.2 million samples.

At the same time, we invested in a Freedom EVO®-HSM workstation for

The Heidelberg CardioBiobank team

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TECAN JOURNAL 2/2016

BIOBANKING

ReliaPrep™ DNA purification*,

Freedom EVO-HSM’s straightforward

also use the excellent 3D simulation tool

taking advantage of the partnership

touchscreen operation guides the

a lot, enabling any potential problems

between Tecan and Promega to

technician through setting up and

such as a mechanical collision to be

implement automated large volume

starting the run, leaving them free

DNA extraction.”

to carry out other tasks while the

identified and resolved before starting

automated extraction is performed.

The step-by-step instructions also mean

The yield and quality of the DNA extracted has significantly improved, which is an unexpected but very welcome benefit.

that less experienced technicians can be taught to use the platform within just a few hours, if necessary, without affecting the results.”

“We currently extract samples from 32 patients per run in about four hours, giving us a throughput of up to 64 samples a day for a normal eight hours operation, with a maximum of 192 extractions per day for 24-hour use. This has increased the speed of extraction by a factor of at least 10,

“Implementation of the system went

perhaps more; the system will pay for

very smoothly. After Tecan had

itself within a short time. In addition, the

completed the installation, a Promega

yield and quality of the DNA extracted

specialist visited the biobank and, in

has significantly improved, which is an

two days, had commissioned the system

unexpected but very welcome benefit.”

and trained four of our staff to use the platform, which is supplied with

“The next bottleneck in our workflow

pre-installed, validated protocols. Unlike

was sample aliquoting. When performed

other suppliers, Promega demonstrates

manually, this is a slow, repetitive

the performance of the DNA extraction during site acceptance testing with a unique biological acceptance test (BAT), using specific, well characterized blood samples to provide a precise, clearly defined reference point. We also validated the system using samples from the biobank and, within a week, were

process with the potential for human errors, and so we have also invested in a Freedom EVO 150 workstation to automate these laborious activities.

the run.”

“There were many reasons for choosing the Tecan workstations, including quality, performance, reliability, durability and robustness in daily operation, and especially the partnership with Promega.

The systems give us the flexibility to adapt our processes as our needs change, futureproofing our procedures, and are also compatible with the new LIMS system due to be implemented, which is crucial. In addition, I have confidence that the systems will be supported for many years, and know that I can communicate directly with local technical experts if I need to, which is invaluable,” Tanja concluded.

To find out more about Tecan’s Freedom EVO-HSM workstation, visit www.tecan.com/reliaprep To learn more about the Heidelberg CardioBiobank, go to www.heidelberg-cardiobiobank. com

We currently run about 20 separate protocols on the platform, giving us the versatility to aliquot different sample

routinely using the workstation. It has

types – including blood, serum, plasma,

been in use every day since then.”

urine and cerebrospinal fluid – for a wide

* For research use only. Not for use in clinical diagnostics.

range of studies, and virtually eliminating “Previously, DNA was manually

human errors. All our technicians find

extracted using standard kits from

it very easy to operate the system, and

a range of providers. Although this

to write specific scripts for individual

worked, it was time consuming and not

studies. The Tube Inspection Unit (TIU)

as effective as the automated process

is a big advantage, helping to enhance

in terms of quality and quantity. The

the performance of the system, and we

TECAN JOURNAL 2/2016

15

IMMUNOLOGY

Building functionality into the antibody supply chain The Monash Antibody Technologies Facility (MATF) in Victoria, Australia, has added a new dimension to its high throughput service, offering screening for antibody functionality.

This time-consuming phase is crucial for many projects, and can now be outsourced to MATF, where comprehensive and flexible automation completes testing in a fraction of the time.

Section of a mouse testis stained with RBM5 antibody (red), binding partner (green) and nuclear stain (blue) (Courtesy of Moira O’Bryan)

16

TECAN JOURNAL 2/2016

IMMUNOLOGY

MATF is a core facility based at Monash University with a global reputation for producing high quality, high affinity monoclonal antibodies for biomedical research projects around the world.

Approximately half of the MATF’s projects stem from the medical and biochemistry faculties at the university, with the remainder from academic institutes, large pharmaceutical and small biotech companies further afield.

Dr Caroline Laverty, Head of Robotics and Manager at MATF, explained: “Our core business is monoclonal antibodies.

We are quite unique with regard to our high throughput capabilities, but our important differentiating factor is delivery of a high quality product.

We are an ISO 9001 certified facility,

Members of the MATF RoboCore team

Without our automation we would need upwards of 20 people to achieve the same output – that’s five times our current head count.

and have a great deal of combined knowledge in the antibody field,

Until recently, MATF has supplied

smooth and very easy to teach. The way

including pharmaceutical industrial

its customers with antibodies that

the deck is configured is ideal for us; we

experience from myself and the facility’s

have been tested by microarray and

can add or take away modules, carriers

director Professor Mark Sleeman. Our

ELISA to confirm that they bind to

or other pieces of equipment very easily.

projects are highly varied, because

the corresponding antigen, at which

And there are other simple, yet really

they all depend on customer-focused,

point the customer has screened

useful things. For example, if an assay

versatile screening strategies that

them in house for functionality. This is

stops halfway through a protocol for any

deliver exactly what the customer

particularly true for the growing field of

reason, it can find itself in space rather

wants. We have tried-and-tested core

therapeutic antibodies. However, this is a

than having to go back to home each

methodologies which we add specific

very time-consuming part of the process

time. The 10 Freedom EVOs® we have

details to, according to what is required

which often takes months. For this

are brilliant – they’re very reliable and

– How many antibodies? What sort of

reason, and at the request of customers,

robust – but the Fluent is something

antigen? How many screening samples?

MATF has invested in a new Fluent®

quite different.”

What is the intended end use? – which is

Laboratory Automation Solution that

where we build in the flexibility.”

can complement this phase of screening.

Caroline concluded: “Our relationship

Caroline explained that the new system

with the Tecan team in Australia is really

“Our robotic systems are very much

will enable them to scale up functional

the icing on the cake, and we actively

integrated into how we find antibodies,

cell-based assays developed internally or

promote this well-built partnership.

and how we screen them. We also

by customers to screen hybridomas. “We

We rely on the service Tecan provides,

operate a smaller liquid handling facility –

will be able to tell them at an early stage

because we have an obligation to our

RoboCore™ – and automation makes the

that the antibodies not only bind, but

customers. We quote timelines and quality

job easier for us and gives us the power

that they are also functional, potentially

and, to achieve that, we need all of our

to be diverse. For every project, we are

saving the customer months, if not

instrumentation to be working all of the

able to generate far higher numbers of

years, of work.”

time. The back-up and application support

hybridomas – and hence a bigger pool

are fundamental for our processes, and

of antibodies for potential screening –

Caroline’s background is in the

I don’t believe that any automation

than would be possible manually. Most

application of automation to varied

company other than Tecan can provide

of the time we’re effectively looking for

laboratory procedures, and she said of

us with that level of assurance.”

a needle in a haystack. The robotics we

the Fluent: “This system is fundamentally

have mean that we can start with a really

different, in ways which are essential

big haystack, and still effectively screen

when you’re trying to deal with so many

it to find the needle. Automation also

To find out more about Tecan’s

projects on the go at the same time. The

gives our staff ‘headspace’ to think

Fluent Laboratory Automation

demographic of our customers’ projects

about the science; it reduces the

is quite wide – these antibodies may be

Solution, visit

amount of staff needed in the lab. We

for anything from veterinary medicine

looked into this a couple of years ago,

to medical research and diagnostics

and without our automation we would

– and we expect to be running up to

need upwards of 20 people to achieve

10 screening campaigns at any one time.

the same output – that’s five times our

The Fluent has been configured to

current head count.”

give us maximum flexibility; it is slick,

www.tecan.com/fluent To learn more about the Monash Antibody Technologies Facility, go to platforms.monash.edu/matf

TECAN JOURNAL 2/2016

17

PROTEOMICS

A more powerful approach to biomarker discovery SomaLogic is using its unique proteomics technology – the SOMAscan® assay – to help identify new biomarkers for a wide range of disease states. Capable of simultaneously measuring thousands of proteins in low volume biological samples, this technique is ideal for the detection of low abundance proteins, perfectly complementing existing technologies for basic science, biomarker discovery and pharmaceutical research.

Biomarker discovery is now at the

allowing precise quantification using

achieve the necessary throughput. It

forefront of medical, diagnostic and drug

standard DNA measurement techniques.

was clear that automation was the way

discovery research, helping to provide

Using our specialized SOMAmer®

forward, making it easier and more

greater insight into many diseases

reagents, this approach enables us to

reliable to perform the assay with fewer

and identify new therapeutic targets.

translate a difficult protein quantification

people. We initially automated the

The size and complexity of the human

problem into a far easier, more

workflow using a liquid handling system

proteome makes the identification of

reproducible and widely accessible

with two 96-channel pipetting arms, but

disease-specific protein biomarkers both

quantitative DNA analysis. Our current

this platform was incompatible with the

difficult and time consuming, requiring

version of the SOMAscan assay is able

SELEX (systematic evolution of ligands

quantitative analysis of thousands of

to measure over 1,300 separate protein

by exponential enrichment) process

individual proteins in complex biological

analytes, over eight logs of dynamic

used to design our SOMAmer reagents,

matrices such as serum, plasma and

range, in just 65 μl of sample, and can

which required independent control

cell lysates. A wide range of proteomic

be performed using various biological

of each pipetting channel. We wanted

methods have been developed to

matrices – including serum, plasma,

to be able to run both workflows on a

offer multiplexed measurement of a

cell supernatants and cell, tissue or

single platform, and chose the Freedom

broad spectrum of protein analytes,

organ lysates.”

EVO® system based on its very high

with mass spectrometry emerging as a front runner. However, many of these techniques offer poor sensitivity for low abundance proteins – requiring extensive depletion steps to access any proteins in the lower end of the dynamic range – have highly limited sample throughput, and reproducibility that disallows detection of smaller signals.

utomation has more than tripled the A throughput of the SOMAscan assay, and the team here is now working with Tecan and our field application specialists to make it even quicker and easier to run.

Colorado-based life sciences company

Throughput is a key consideration for

flexibility and the ease with which we

SomaLogic has developed a unique

any biomarker discovery workflow,

could configure the instrument for

technology allowing the measurement of

particularly for the pharmaceutical

different processes.”

a broad range of protein concentrations

sector, requiring effective and

in small sample volumes, complementing

reproducible liquid handling to

“We selected the Freedom EVO 200

existing techniques and providing

ensure accurate results. Dr Stephan

because of the large number of reagents,

improved measurement of low

Kraemer, Director of Assay Execution,

assay plates and devices – including

abundance proteins. Dr Tim Bauer,

commented: “When we first developed

two Te-VacS™ modules and four shakers

Associate Director at SomaLogic,

the SOMAscan assay in a microplate

(Q.Instruments) – which needed to be

explained: “Our technology – the

format, we were running it manually.

fitted onto the workdeck,” Stephan

SOMAscan assay – converts a protein

Three technicians needed to stand

continued. “The system also has both

concentration into a DNA concentration,

side-by-side, pipetting in parallel to

an eight-channel Liquid Handling Arm,

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TECAN JOURNAL 2/2016

PROTEOMICS

To find out more about Tecan’s protein science solutions, visit www.tecan.com/proteinscience To learn more about SomaLogic, go to www.somalogic.com

Dr Stephan Kraemer with one of SomaLogic’s Freedom EVO platforms

allowing independent control of

the logistical challenges of shipping

individual pipetting channels, and

their samples to us for analysis. The

a MultiChannel Arm™ (MCA) for

use of standardized protocols should

simultaneous pipetting into all 96 wells

also allow direct comparisons of results

of an assay plate. This gives us the

between sites.”

flexibility to perform either the SOMAscan assay or the SELEX target

“We have only recently placed the

identification workflow on the same

first instruments with customers,

instrument, or to run both together.”

but the feedback we have received so far has been very positive. Automation

Tim added: “Since introducing our

has more than tripled the throughput

first Freedom EVO platform, we

of the SOMAscan assay, and the team

have purchased three additional,

here is now working with Tecan and

almost identical systems, and we are

our field application specialists to

very happy with both the precision

make it even quicker and easier to run,”

and robustness of the instruments.

Stephan concluded.

Now that we have established our automated workflows in house, we are working with Tecan to optimize and place instruments using the same configuration at customer sites around the world, initially focusing on human serum and plasma samples.

This will allow customers to run more samples much more quickly, without

TECAN JOURNAL 2/2016

19

CLINICAL DIAGNOSTICS

Consistency for a changing market PerkinElmer Healthcare Diagnostics (Shanghai) Co develops and manufactures instruments for the processing and analysis of clinical samples in hospital laboratories across China. A pioneer of time-resolved fluorescence techniques in this market, the company prides itself on creating cutting-edge automated solutions to address unmet needs in infectious disease diagnostics.

The in vitro diagnostics (IVD) market

who understand the demands of the

facility develops and manufactures a

has very specific needs when it comes

diagnostics sector, ensuring ongoing

range of Sym-Bio brand IVD instruments

to laboratory automation, requiring

support and allowing the development

for use in hospitals across China,

highly integrated solutions that are

of new solutions to address an ever-

focusing primarily on the detection

both robust and easy to use. Unlike the

changing market.

of infectious diseases. Combining

life sciences research sector – where

our proprietary TRF immunoassay

flexibility and adaptability are often

PerkinElmer Healthcare Diagnostics

technologies with advanced laboratory

essential to guarantee productivity

(Shanghai) Co – formerly Shanghai

automation, our aim is to provide

as project needs change – automated

Sym-Bio Life Science Co – is a leading

convenient and cost-effective testing

systems for clinical laboratories should

provider of diagnostics instruments and

for a range of diseases, many of which

offer highly reproducible results

related reagents for Chinese hospitals.

are not served by other solutions on

over several years, while requiring

Originally founded to develop and

the market.”

only minimal maintenance and user

commercialize novel time-resolved

expertise. The use of high quality OEM

fluorescence (TRF) techniques, the

When developing its EasyCuta 1260

components is therefore key to the

company is now a major innovator in

automatic time-resolved fluorescence

long-term success of IVD instruments,

the local IVD market, with a dedicated

analyzer, the company chose to partner

providing consistent performance over

R&D center in Taicang, Jiangsu Province.

with Tecan, taking advantage of the

time. However, it is just as important to

Torres Cai, Instrument Sourcing

company’s liquid handling expertise

work with knowledgeable OEM partners

Supervisor, commented: “Our Taicang

and strong local presence to ensure the success of the project. Cai explained: “Precise transfer of samples and reagents is crucial for the accuracy and reproducibility of immunoassay results, making syringe pumps the most important component in the EasyCuta 1260. When selecting an OEM supplier, our product developers looked not only for the best technical solution, but also for a very high level of technical and application support. Staff from Tecan’s local organization, based in nearby Shanghai, were very responsive and knowledgeable, giving us confidence in both the solutions and support on offer.”

“After discussing the needs of our instrument with the Tecan team, we chose the Cavro® XLP syringe pump for the EasyCuta 1260 project.

Tecan Cavro components are essential to the consistent and reliable operation of the

Each instrument contains six individual

EasyCuta instruments

XLP pumps which are required to

20

TECAN JOURNAL 2/2016

CLINICAL DIAGNOSTICS

perform advanced liquid handling reliably for long periods, so it was crucial that our engineers understood how to maximize long-term performance.

We received very good support from Tecan during instrument development and validation; the company provided periodic training for our engineers to

Precise transfer of samples and reagents is crucial for the accuracy and reproducibility of immunoassay results.

ensure we could optimize instrument performance and provide appropriate maintenance schedules, and its application experts were always available to answer any questions we had. Since its launch in 2013, we have installed EasyCuta 1260 platforms in over 400 hospitals, and we have been very happy with the performance of both the instrument and the Cavro pumps.”

In 2014, PerkinElmer began developing a second system – the EasyCuta MiNi – to provide a lower throughput solution for smaller hospitals. “Based on feedback from existing EasyCuta 1260 customers,

To find out more about Tecan

we took this opportunity to enhance

Cavro components, visit

the instrument workflow and shorten

www.tecan.com/components

turnaround times,” Cai continued. “In addition to modifying the assay method slightly, we chose to use the Cavro ADP (Air Displacement Pipettor) to allow faster liquid transfers, helping to reduce the overall processing time. Working with Tecan made it very straightforward

To learn more about PerkinElmer Healthcare Diagnostics (Shanghai), go to www.perkinelmer.com.cn/ products/default.xhtml

to transfer our method from syringe pumps to pipettor technology, and this is one of the benefits of working with a leading OEM supplier with a diverse product portfolio.”

TECAN JOURNAL 2/2016

21

CELL BIOLOGY

A modern elixir of life Scientists at Tokyo University of Technology are searching for foodstuffs that can slow the aging process. Focusing on mitochondrial regulation, researchers from the university’s School of Bioscience and Biotechnology are using cell-based assays to help identify food components with anti-aging properties.

Since time immemorial, people have

production, reducing the amount of

time-course assays. “The ability to

dreamt of discovering ways of holding

food required and limiting the stress on

perform measurements that are

back the aging process. A group of

the body. As many foods contain these

both quantitative and time-based

researchers in Japan are seeking to

organic acids, it is possible to consume

is a great advantage. Previously,

turn this long-held dream into reality,

them as part of a normal diet.”

it was not possible to satisfy both needs simultaneously; it had to

looking at the anti-aging properties of specific foodstuffs. This research

Professor Satoh and his team are

be one or the other. Quantitative

focuses on the effect of various food

screening for compounds that are able

fluorescence measurements made

components on mitochondria, which

to promote mitochondrial activity using

using a conventional microplate reader

are believed to play an important role in determining the survival or death of the cells and are more strongly influenced by reactive oxygen species (ROS) than other organelles. Professor Takumi Satoh, Head of the Anti-aging Food

aving this quality and breadth of data H makes a real difference when preparing manuscripts for publication.

Laboratory, Advanced Food Course at Tokyo University of Technology’s

microplate-based cellular assays, aided

give satisfactory one-off readings,

School of Bioscience and Biotechnology,

by a Spark® 10M multimode microplate

but there is no capability to make

explained: “I believe that the ability

reader. He continued: “We culture

repeat measurements over time

of food components to activate

adherent cells in microplates and, using

for cell-based assays. Conversely,

mitochondria is important for anti-aging.

a fluorescent dye, we can visualize

fluorescence microscopy is well suited

Consequently, we are searching for

changes in mitochondrial membrane

to measurements over extended time

compounds in food that can enter and

potential and monitor intracellular

periods, but this method can only

stimulate mitochondria directly. We are

ROS levels caused by test compounds.

collect data from a single cell. The

currently investigating various organic

We have initially been working on the

Spark 10M was therefore an important

acids, such as 3-hydroxybutyric acid,

establishment of this method using the

breakthrough for our work. We now

which is from a group of compounds

Spark reader, and have already started

have a microplate reader that makes

known as ketone bodies that have been

to see some exciting results.”

it possible to simultaneously assay a population of cells – measuring

reported to prolong the lifespan of C. elegans. These are relatively small

The Spark reader’s integrated Gas

ROS concentrations or mitochondrial

compounds and, unlike compounds

Control Module (GCM™) is capable

membrane potentials – both

such as glucose, some can enter the

of precise regulation of temperature

quantitatively and temporally, allowing

mitochondria directly without the need

and carbon dioxide concentration,

us to determine everything in one go.

for prior catalysis in the cytoplasm.

which maintains cell health and

We can also run multiple experiments

This results in very efficient energy

allows researchers to perform longer

at once, usually in 24-well microplates.”

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TECAN JOURNAL 2/2016

CELL BIOLOGY

“It is a great benefit for our work to be able to ensure that the cells are alive and healthy for the duration of the assay. Both the temperature and the carbon dioxide concentration can be tightly controlled, allowing complete assays to be carried out under the same conditions as within a cell culture incubator. Until now, this combination of features was not available, so the length of time during which measurements could be taken was very limited.”

“Another really good thing is that we can run assays continuously for many hours.

A common problem with fluorescence microscopy is the non-specific

Professor Satoh and student Zhang Jian with the Spark reader

background effect, which increases with prolonged exposure time. However, with the Spark 10M, the measurements are made very quickly, minimizing background interference and allowing us to collect data over much longer periods.

Having this quality and breadth of data makes a real difference when preparing

To find out more about Tecan’s Spark 10M reader, visit www.tecan.com/spark10m To learn more about the

manuscripts for publication. Moreover,

Tokyo University of Technology’s

its operation is simple, making it very

School of Bioscience and

easy to use, especially when compared

Biotechnology, visit

to some other instruments I have seen.

www.teu.ac.jp/english/

It is very good for our experiments,”

departments/010614.html

Dr Satoh concluded.

TECAN JOURNAL 2/2016

23

PROTEIN SCIENCE

Miniaturizing separation science The development and characterization of new chromatography media is both complex and time consuming. At Merck KGaA in Darmstadt, Germany, the implementation of automated miniature column chromatography protocols has helped to improve throughput and reduce the amount of material required, allowing more measurements to be performed in less time.

Protein purification is a crucial step in

must be performed to characterize the

development, and within six months

the production of biopharmaceuticals,

different candidates, including the

had successfully developed and

and is frequently performed using

determination of selectivity, and static

validated our automated miniature

chromatographic processes. To help

and dynamic binding capacities. The

column chromatography method. We

biotech companies accelerate their

dynamic binding capacity is determined

can screen eight columns in parallel,

product development, manufacturers

under flow conditions, and was

completing dynamic binding capacity

are continually striving to develop new

previously carried out using standard 1,

determinations and resin selectivity

chromatography media, screening

5 or 10 ml lab-scale columns. However,

measurements in three to six hours,

prototype resins with novel functionalities

a large amount of feed material was

depending on the operating

and improved performance to extend

required for each column, and

parameters, compared to over 24 hours

their product portfolio.

evaluation of all the different

for sequential testing on a conventional

candidates was very time consuming.

system. We use 5- to 10-fold less feed

The Chromatography R&D and

We needed to reduce the feed amount

material, and the labor intensity is

Manufacturing Science and Technology

and perform more measurements in

reduced, freeing up more time for us

(MSAT) groups of Merck’s life science

less time, and this led us towards

to perform other valuable tasks.”

business are both engaged in the

miniature column chromatography.”

development of specialist

Senior Applications Engineer Andreas

chromatographic media – such as

André Kiesewetter, an MSAT

Stein added: “We received valuable

Fractogel®, Eshmuno® and ProSep® – for

Applications Engineer, takes up the

support from Tecan during the

the effective and efficient purification

story: “We decided to upgrade our

implementation period, and also

of biomolecules, as well as the creation

in-house robotic capabilities to include

benefitted from the availability of an

of supporting resin performance

state-of-the-art automated miniature

application note describing the optimum

application data. The groups actively

column chromatography using Atoll

performance settings for automated

support the company’s worldwide

RoboColumns®, a proven, user-friendly

miniature column chromatography.

customer base through a network of

technology that can be easily

Programming the workstation is easy

scientists and engineers, as well as

implemented on a liquid handling

to learn, and the integration between

10 M Lab™ collaboration centers

platform. We compared various

Freedom EVOware® and other programs,

located across the globe. The M Lab

workstations, eventually deciding that

such as visual basic scripts, is a huge

collaboration centers in Europe, Asia,

the close collaboration between Atoll

advantage; a system with an open

and the US are equipped with similar

and Tecan – and the popularity of the

program is much more flexible.”

liquid handling platforms configured

technology – made the combination of

for miniature column chromatography,

RoboColumns and the Freedom EVO®

“Automated high throughput

ensuring that customers receive the

workstation the ideal solution for both

experiments generate many samples for

same level of support regardless of

resin and application development

analysis, which could potentially create

location, as well as simplifying method

assays. We can also take advantage of

a bottleneck. Now that the miniature

transfer between sites for flexible

the system’s liquid handling capabilities

column chromatography application is

allocation of resources. Peter Menstell,

to perform dilutions and other sample

running really well, our aim is to go a

Applications Engineer from the

preparation steps in our workflows.”

step further and automate the analytics, establishing a fully integrated workflow

Chromatography R&D group, explained: “When a new resin is under

André continued: “We ran the first trials

to alleviate any possible hold-ups,”

development, a variety of experiments

after just a few weeks of protocol

concluded Andreas.

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TECAN JOURNAL 2/2016

PROTEIN SCIENCE

We use 5- to 10-fold less feed material, and the labor intensity is reduced, freeing up more time for us to perform other valuable tasks.

To find out more about Tecan’s protein science solutions, visit www.tecan.com/proteinscience To learn more about Merck’s life science business, go to www.merckmillipore.com

From left to right, Peter Menstell, André Kiesewetter and Andreas Stein

TECAN JOURNAL 2/2016

25

CLINICAL DIAGNOSTICS

Automation – an extension of the analyst Reputation and accreditation are essential for clinical reference laboratories, helping to win new business and ensure a steady flow of work from existing customer labs.

With diagnostic testing demands increasing year-on-year, Carpermor continually invests in cutting-edge assays and state-of-the-art laboratory automation to maintain the quality and efficiency of its services.

all diagnostic disciplines, and quality is

in question. They allow us to process

the most important aspect of our work;

the necessary volume of samples in a

we are accredited by both the College

reproducible and reliable manner, and

of American Pathologists (CAP) and

we do not have any problems with the

our national accreditation authority,

accuracy or repeatability of our ELISAs,

Entidad Mexicana de Acreditación.”

which has helped with our overall workflow by minimizing the need for

Mario continued: “With such a large

repeat testing. As with any technology,

workload and just 185 staff, laboratory

there was a learning period where we

automation is now crucial to ensure

needed to ‘tame’ the equipment to our

that we maintain the level of service

needs, but the Tecan platforms have

our customers expect, in terms of

been excellent since we completed this

both quality and turnaround times.

process. We effectively think of the

Carpermor is an international

Previously, we worked entirely

instruments as working prostheses – an

reference laboratory in Mexico City,

manually – with various individual

extension of the analyst. The flexibility

offering a full range of routine and

washers, readers, etc. – but the rapidly

of the equipment is another advantage,

specialist clinical diagnostic services

increasing number and diversity of

allowing us to perform a wide range

to healthcare providers across Mexico,

tests we needed to perform made this

of protocols on a single platform, and

Carpermor’s Elena Alcántara and Mario Garcia

this has helped us to move away from

Laboratory automation is now crucial to ensure that we maintain the level of service our customers expect.

isotopic analyses for many analytes.”

as well as participating in contract

unsustainable. Our main requirement

we have been trying to replace all our

and academic research projects

for any automated equipment within

assays that rely on isotope analysis

globally. The laboratory prides itself

the lab is that it is extremely reliable,

with either chemiluminescence- or

on its exceptional levels of customer

as our throughput means that even a

ELISA-based methods. Our relationship

satisfaction, using state-of-the-art

short period of downtime can lead to

with IBL International has been crucial

technology to ensure a fast and

a significant backlog. We also need

to achieving this, and we have been

efficient service for its 1,500 customer

equipment that is very versatile, helping

able to work closely with both the

labs. Mario Garcia, General Manager for

us to cope with a constantly changing

company and its distributor here in

Carpermor’s Laboratory and Analytical

portfolio of assays. For these reasons,

Mexico to achieve our goals. Although

Centers, explained their approach: “We

we chose Tecan’s Freedom EVOlyzer®

we are not generally well served by

strive towards service excellence and

and Freedom EVO® platforms to

major diagnostics providers in Latin

exceptional accuracy, providing a high

perform the bulk of our ELISA testing.”

America – they are quite inflexible

Elena Alcántara, a member of the laboratory team at Carpermor, explained: “Isotopic techniques are both complex and expensive, and so

quality, efficient and competitively

to our local needs and you have to

priced service for both routine and

“We have worked with Tecan for a

do a lot of the work yourselves – we

highly specialized testing. We perform

number of years, and the quality of

have found that this was not the case

over 32 million analyses a year across

these workstations has never been

with IBL International. The company’s

26

TECAN JOURNAL 2/2016

CLINICAL DIAGNOSTICS

Each platform offers the flexibility to run

Carperpor’s workflow relies on Freedom EVOlyzer and Freedom EVO platforms to automate

multiple protocols, working as an extension of

ELISA testing

the analyst

attitude is very refreshing; we have

kits and the performance of our Tecan

always found them to be very flexible

platforms. Our ELISA portfolio is still

to our requirements, and we get an

growing, and we will soon need another

answer to any queries we have within

Freedom EVOlyzer, so we hope to

24 hours. They are certainly in tune

continue with Tecan and IBL assays for

with our needs.”

many years to come,” Elena concluded.

“The IBL assay kits themselves are also very good. They are all fully validated,

To find out more about the IBL

which is obviously important in an

range of diagnostic assays, visit

accredited lab, and the convenience

www.tecan.com/immunoassays

of the infectious disease assays has helped us to resolve a bottleneck in our workflow. The expiry characteristics of the assay reagents are another important consideration, particularly for low volume tests that we do not perform regularly. The IBL kits are very stable,

To find out more about Tecan’s diagnostic automation portfolio, visit diagnostics.tecan.com To learn more about

with long expiry dates, which simplifies

Carpermor’s services, go to

logistics and ensures we can meet our

www.carpermor.com.mx

turnaround time targets, even for tests that we rarely perform. Overall, we are very happy with both the IBL ELISA

TECAN JOURNAL 2/2016

27

3D CELL CULTURE

A 3D revolution A novel 3D cell culture platform is enabling scientists to generate microtissues that closely mimic the behavior of human tissues from virtually any cell type. This proprietary technology is easily automated, offering speed and versatility, combined with consistency and reproducibility, for high throughput applications.

Classical 2D cell culture is a well-proven

or days required by other methods –

Chris continued: “Scaffold-free methods

technique widely used in biological

and is easily automated on a liquid

are also extremely dependent on cell

assays, drug screening and many other

handling workstation. Chris Millan,

type and donor, and may not always be

research areas. However, cells grown in

Chief Technical Officer at CellSpring,

able to form 3D structures, whereas 3D

a monolayer behave very differently to

explained: “3D Bloom offers several

Bloom can incorporate virtually any cell

cells in a real tissue, and so the response

advantages compared to other

type into the system. A further restriction

of 2D cultured cells to drugs under

methods of creating 3D microtissues.

of the spheroid technique is that it is not

development is not an ideal indicator

Scaffold-free approaches – spheroids

generally possible to have more than

of the likely in vivo response. The 3D

and hanging-drops – for example, take

10,000 cells per microtissue. This limits

Bloom® Biopolymer Platform from

three to five days to generate a 3D

the downstream readout, as it is difficult

CellSpring, an ETH Zurich spin-out,

structure. With 3D Bloom, two

to generate enough DNA or protein to

is showing promise for improving

naturally-derived biopolymers are

establish what is happening with those

the predictive capabilities of in vitro

sequentially pipetted into a standard

cells. In contrast, 3D Bloom is viable

preclinical testing for drug safety and

multiwell plate, where a spontaneous,

over a very wide range – from 5,000 to

efficacy. This proprietary technology

cell-friendly cross-linking reaction

2 million cells per microtissue – removing

can generate 3D microtissues in just

assembles the cells into a 3D structure

this limitation. Variability is another

30 minutes – far less than the hours

in as little as 30 minutes.”

potential issue; scaffold-based processes use materials isolated from animals and consequently may vary between batches by as much as 50 %. Our technology, being very chemically defined, offers almost 100 % reproducibility.”

“At the moment, our focus is on high throughput testing of new cancer drugs, looking specifically at the expression of targets of interest to pharmaceutical companies, which are generally over expressed by cells cultured in our system. As a result, the sensitivity of the system to candidate drugs is higher than with conventional methods, which is an advantage when you’re trying to compare numerous different formulations to determine the best candidates,” said Chris. “Performing this work manually is both time consuming and operator dependent, and so automation is clearly the way forward. Through the ETH Innovation and Entrepreneurship Lab (ieLab), we made contact with Tecan, and have worked closely with the company since then to automate our technology on a Freedom EVO® platform.”

3D Bloom allows rapid, automated generation of 3D cell cultures

28

TECAN JOURNAL 2/2016

3D CELL CULTURE

“Initially, Tecan’s specialists worked side by side with us, showing us how to program the system and automate our 3D cell culture method. The Freedom EVO proved extremely straightforward to operate, and we can easily adjust the scripts ourselves if necessary. It’s been incredible; speed and reproducibility have improved tremendously. We can now reliably process 10 plates in the time it takes to run a single 96-well plate manually, and staff are free to carry out other tasks. In the next phase of the collaboration, we plan to investigate co-culture models, incorporating different cell types into a single microtissue. This cannot be performed manually – you do not have enough control when positioning small droplets by eye – but the liquid handling platform should be able to accomplish it quite easily. It’s very apparent to us that we will be selling our products

CellSpring’s Kramer Schmit (CEO, left) and Chris Millan (CTO, right) are hoping the partnership with Tecan will expand the potential of the 3D Bloom platform for screening applications

exclusively for automated platforms, and particularly the Freedom EVO, which will also form the basis of our in-house service offering. There is no doubt that automation has helped us get to market much faster than would otherwise have

It’s been incredible; the speed and reproducibility have improved tremendously.

been possible, and the collaboration has gone so well that we are now entering into a co-marketing agreement with Tecan,” Chris concluded.

To find out more about Tecan’s 3D cell culture solutions, visit www.tecan.com/3dcellculture To learn more about CellSpring, go to www.cellspring.co

TECAN JOURNAL 2/2016

29

IMMUNOLOGY

HMGB1 – a silent partner in cancer?

Many people in the life science and medical communities will never have heard of HMGB1 or, to give it its full title, high mobility group box 1 protein. Despite this, it is the most common nuclear protein after the histones – with around a million copies per cell – and plays a key role in the body’s response to everything from trauma and infection to a stroke and heart attack. Dr Michael T Lotze, Professor of Surgery, Immunology and Bioengineering at the University of Pittsburgh School of Medicine, discusses the clinical interest in this remarkable protein for the diagnosis and treatment of patients with cancer.

Dr Michael T Lotze, University of Pittsburgh School of Medicine

Cell death in animals normally occurs via a highly regulated, carefully controlled process known as apoptosis. Designed to avoid the cell contents spilling out into the surrounding tissue and causing damage to neighboring cells, it results in the formation of carefully packaged apoptotic bodies which can be quickly and easily engulfed by phagocytic immune cells. In contrast to this, cancer cells do not undergo a programmed cell death, instead experiencing the unscheduled necrotic cell death normally associated with cellular trauma.

Necrosis results in the loss of membrane integrity, leading to the release of the cellular carcass material into the extracellular space.

A clinical interest One of the proteins released into tissues and the blood by the death of cancerous cells is HMGB1, a chromatin protein normally associated with the packaging of DNA in the cellular nucleus. In most animals, the presence of HMGB1 in the extracellular space is a critical ‘alarm signal’ – known as a damage-associated molecular pattern (DAMP) – triggering an inflammatory immune response to repair the damage. It also affects the surrounding cells directly by inducing autophagy, a ‘self-preservation’ mechanism which helps the intact cells to survive and repair themselves. Under normal circumstances, this would be 30

TECAN JOURNAL 2/2016

IMMUNOLOGY

beneficial, but when the cell death is a result of chemotherapy, and the target cell that is being instructed to survive is a tumor cell, then you obviously have a problem. This not only promotes survival of the tumor cells, but it can also reduce their susceptibility to chemotherapy drugs. In addition, the localized presence of HMGB1 can suppress immunity, hindering natural antitumor responses such as T cell activation.

As a result, there is now significant interest in inhibiting HMGB1 in cancer patients – either directly, or up- or downstream – to suppress autophagy and the ‘self-preservation’ response in tumor cells. Early studies combining traditional chemotherapy with HMGB1-targeting immunotherapy have been remarkably successful in patients with pancreatic and kidney cancers, and additional trials are now being launched.

HMGB1 isn’t only a problem at the tumor site either. Effective chemotherapy can lead to a significant and sustained rise in circulating HMGB1 – as well as other intracellular molecules, such as uric acid, potassium and nucleic acids – as the tumor responds to treatment.

When released into the bloodstream,

HMGB1 is a key mediator for the immune system, signaling cellular stress, and can play a vital role as a biomarker for arthritis, stroke, cancer and sepsis.

these molecules are filtered out by the kidneys, leading to a serious, and potentially fatal, form of kidney damage

other approaches – nothing else comes

known as tumor lysis syndrome.

close – and the company also offers

Scan the

a range of complimentary products,

QR code to

We know that HMGB1 is a critically important molecule, not just in cancer, but also in… many other biological processes.

Furthering research One of the major challenges of HMGB1 is the availability of appropriate tools for

such as the Anti-HMGB1 Chicken IgY Neutralizing Polyclonal Antibody, making it faster and easier to conduct our

download the poster

investigations. We know that HMGB1 is a critically important molecule, not just in cancer, but also in autoimmunity, trauma, infection, myocardial infarction, stroke and many other biological processes that are not yet fully understood. It’s becoming increasingly clear that it is

To find out more about IBL’s HMGB1 ELISA kits, visit www.tecan.com/immunoassays

important throughout medicine, and having the right tools available will help

To learn more about

us to further our understanding and

Dr Lotze’s research, go to

improve clinical approaches in the future.

www.immunology.pitt.edu/ person/michael-lotze-md

research. Although many immunology and biochemistry groups around the world have attempted to develop ELISAs for the accurate quantification of HMGB1, it wasn’t until the launch of IBL International’s HMGB1 ELISA* that a reliable method was available. This assay system is intrinsically more effective than

The IBL HMGB1 ELISA Kit is CE marked for IVD use in Europe, but is for research use only in the USA. *Developed by Shino-Test Corporation, Japan, in co-operation with Prof Dr Ikuro Maruyama of Kagoshima University, this product is exclusively distributed worldwide

by IBL International, except in Japan, China (incl. Hong Kong) and Taiwan.

TECAN JOURNAL 2/2016

31

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