PharmaNEWS - European Pharmaceutical Review

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PharmaNEWS - European Pharmaceutical Review

DIGITAL

ISSUE 01 2010

TECHNOLOGY • KNOWLEDGE • INNOVATION

PharmaNEWS

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Lab Automation series


Helen Difford

Editor

European Pharmaceutical Review

email: hdifford@russellpublishing.com

Welcome to the first issue of

European Pharmaceutical Review

Digital 2010!

Throughout the year, we will be featuring a second series of articles

from Dr Sheraz Gul on lab automation. In this issue, Dr Gul looks at an

overview of the common types of assays and emerging trends for the

protease enzyme target class. You can find his article on page …

Don’t forget to check out Dr Gul’s previous lab automation series in

past issues of European Pharmaceutical Review

Digital, found on

www.europeanpharmaceuticalreview.com.

As always, if you would like to contribute to a

future issue of EPRDigital by providing editorial,

news or include your latest press

release, please do not hesitate to

contact me.

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PharmaNEWS

A REGULAR ROUND-UP OF INDUSTRY PRODUCTS, INNOVATION, NEWS AND DEVELOPMENTS

DropSense96

3 PharmaNEWS

The Trinean DropSense96® is a new multichannel spectrophotometer for quick and precise UV-VIS spectral

analysis of microliter droplets of DNA/RNA, protein or small compounds. This spectrophotometer has all

the characteristics of a plate reader, but uses specific DropPlate 16/96 chips with 1-2,5 μl samples.

The unique features of the DropSense96 are






UV-VIS absorption spectrometry

Full spectrum analysis of 96 droplet samples in less than 5 minutes

Compatible with both DropPlate 16/96 and classic 96well microplates

Predefined protocols for DNA, RNA and protein concentration measurements

User-friendly software on a remote PC


ISSUE 1 2010

4 PharmaNEWS

Need an easy way to cleanup PCR

products? Without using a column or

beads? And without losing product?

USB® ExoSAP-IT® is the solution.

Single step PCR cleanup

■ Add ExoSAP-IT reagent directly to PCR product

Eliminates spin columns

■ Decreases time and expense while increasing yield

No sample loss

■ 100% recovery of both short and long PCR products

Removes contaminating primers & dNTPs

■ No interference in downstream applications

Simple processing

■ Lends itself to robotics

■ Replaces beads, filtrations and plates

Scaleable

■ Economical for high throughput purification

ExoSAP-IT reagent

ExoSAP-IT reagent is designed for simple, quick PCR cleanup for

downstream applications, such as sequencing or SNP analysis.

When PCR amplification is complete, any unconsumed dNTPs

and primers remaining in the PCR product mixture will interfere

with these methods.

ExoSAP-IT cleanup reagent is added directly to the PCR product

and incubated at 37°C for 15 min. The enzymes are active in the

buffer used for PCR, so no buffer exchange is required.

After PCR treatment, ExoSAP-IT reagent is inactivated simply by

heating to 80°C for 15 min.

The ExoSAP-IT method utilizes two hydrolytic enzymes,

Exonuclease I and Shrimp Alkaline Phosphatase, together in a

specially formulated buffer, to remove unwanted dNTPs and

primers from PCR products. Exonuclease I removes residual

single-stranded primers and any extraneous single-stranded DNA

produced in the PCR. Shrimp Alkaline Phosphatase

removes the remaining dNTPs from the PCR mixture which

would interfere with the subsequent reactions.

www.europeanpharmaceuticalreview.com


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Pall Corporation and SOTAX deliver costeffective

automation certified solutions for

automated pharmaceutical testing systems

Pall Corporation, (NYSE: PLL), a global leader in

filtration, separation and purification, has announced

a long-term joint marketing agreement with SOTAX

Group that will include Pall Acrodisc® PSF syringe

filters in all SOTAX dissolution systems for solid

dosage forms. The exclusive agreement is the initial

step that allows the companies to begin delivering

complete certification solutions that save time and

testing costs for quality control laboratories.

Pall Acrodisc PSF syringe filters are used by

research and analytical laboratories to help expedite

automated sample processing. First introduced in

1973, Pall’s Acrodisc syringe filters provide

improved retention efficiency to help extend HPLC

column life by as much as 46 times. The multilayered

prefilter provides two to four times the

throughput of standard glass fiber prefilter devices

to allow for quick and easy filtration of samples that

are difficult to filter. The PSF housing has been

designed for smooth operation in automated systems

and operating pressures, eliminating filter burst.

SOTAX is a Swiss-based provider of highquality

dissolution testing systems, composite assay

and content uniformity workstations along with

physical tablet testing instruments for the

pharmaceutical industry. Dissolution testing is used

by the pharmaceutical industry to characterize the

dissolution properties of an active drug, an active

drug's release and the dissolution from a dosage

formulation. SOTAX systems formulate the drug

dosage form and develop quality control

specifications for the manufacturing process.

“By exclusively recommending Pall Acrodisc

PSF technology as part of our testing systems, we

can ensure we are providing the pharmaceutical

industry with a significantly improved automated

dissolution process for multiple unattended tests.

This includes applications for media preparation and

delivery, automated dosage introduction, automated

sampling, filtering and sample analysis,” said Gilles

Devidts, Head of Product Management, SOTAX.

At the start of this agreement, all new SOTAX

AT 70smart Dissolution systems and SOTAX CTS

Content Uniformity test systems will include the

Pall Acrodisc PSF syringe filters. Filter refills will

be available directly from Pall Life Sciences

through its regular sales and distribution channel.

“This marketing partnership with SOTAX

advances the performance of the automated

laboratory analysis platform,” said Larry O’Connell,

Senior Vice President, Global Lab Products for Pall

Life Sciences. “Customers can now obtain the dual

benefits associated with SOTAX and Pall

technology to achieve superior analysis and

performance.”

To learn more about Acrodisc PSF syringe filters,

visit www.pall.com/lab.

5 PharmaNEWS


ISSUE 1 2010

Bio-Rad’s CFX automation system enables

walk-away, high-throughput real-time

6 PharmaNEWS

In the pharmaceutical and biopharmaceutical markets, performance, time-to-result and sample throughput

are increasingly important. The addition of Bio-Rad’s CFX automation system to the CFX96 and CFX384

real-time PCR systems offers a high quality, high-throughput, automated solution for these researchers.

“The CFX automated system is targeted at anyone who wants to increase their throughput and run their

real-time PCR system in an automated environment,” said Richard Kurtz, Senior Marketing Manager, Bio-

Rad Laboratories.

According to Kurtz, the primary benefit of the system is its ability to enable hands-off operation of the

CFX96 and CFX384 instruments. “It eliminates the bottleneck of needing to have someone present to run

the experiments. Now you can run your experiments overnight without having to be present to put plates

into the system,” he said.

The CFX automation system is equipped with a bench-top plate handler with the capacity to load up to

twenty 384-well plates or 7,680 samples at a time on the CFX384 system, facilitating workflow

automation, generation of large volumes of data and rapid analysis of that data.

Researchers can use the CFX automation control software to configure their batch run of plates,

selecting to run in a quick start mode with the same protocol for all runs, or in an advanced mode enabling

different protocols for each plate.

Upon run completion, users can analyse data when and where they want by choosing to be sent an email

notification with an attached data file. They can also request an automatically generated PDF report in

addition to the data file.

www.bio-rad.com/amplification

Integromics and TATAA Biocenter collaborate

to offer comprehensive qPCR data analysis

Integromics S.L., the leading provider of

bioinformatics solutions to gene expression

researchers, has announced an exciting collaboration

with the TATAA Biocenters in Sweden and the

Czech Republic to provide expert training in qPCR

data analysis using RealTime StatMiner. Professor

Mikael Kubista, Chairman of TATAA Biocenter,

explained: “We have been teaching courses in realtime

qPCR since 2001, and continually update our

course programs to reflect the latest technological

advances. Our data analysis courses are designed for

researchers, technicians and post-graduate students

without specialist bioinformatics training, and

RealTime StatMiner provides these users with an

array of powerful, easy-to-use data analysis tools.”

Imad Yassin, Commercial Director at Integromics,

added: “Our sophisticated and powerful software

packages are designed to provide customers with all

the features they need, in a user-friendly format. The

intuitive interface and unique ‘guided workflow’

approach of RealTime StatMiner provide an

unparalleled solution for in-house analysis of qPCR

and microarray data, offering step-by-step guidance

for straightforward analysis and interpretation. We

are excited that the TATAA Biocenter – a respected

leader in PCR services and training – has recognised

the value of RealTime StatMiner, and wishes to

share the benefits of this comprehensive package

with its customers.”

To find out more about Integromics RealTime

StatMiner, visit

www.integromics.com/StatMiner

To find out more about TATAA Biocenter

training in data mining visit:

http://www.tataa.com/Courses/Courses.html

Meet Integromics and TATAA at qPCR

www.europeanpharmaceuticalreview.com


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Astech Projects expands its packaging

robotics and automation offering

Leading automation systems developer Astech Projects has announced the appointment of John Roberts to

expand its dedicated sales team within the Packaging Robotics and Consumer Goods Testing markets. Mr

Roberts assumes the post of Business Development Manager, adding to Astech’s team of high calibre

applications engineers.

With over 15 years of experience in the packaging industry, Mr Roberts has built up a vast array of

industry supplier/customer relationships. Astech are hoping to utilise his extensive experience and firmly

establish Astech as a world class systems integrator within these highly specialised markets.

Anthony Moran, Director, Astech Projects, said “We are delighted to welcome John to the team.

Recruiting him is part of a strategic strengthening of Astech’s offering for the packaging industry. He is

highly regarded and respected within the industry and I am looking forward to having his expertise as

Astech continues to expand.”

www.astechprojects.com

Concentrating environmental samples

prior to analysis

For environmental laboratories, it is of great benefit to concentrate samples prior to analysis to help

accurately determine levels of analyte in a given sample.

Genevac's proprietary SampleGenie flask technology has been developed to help scientists eliminate

handling of samples by concentrating samples, without loss of volatile components, directly into a small

GC or HPLC vial.

SampleGenie technology enables samples in Genevac ROCKET, EZ-2 or HT-series centrifugal

evaporators to be concentrated directly into a single vial, eliminating the need for reformatting of samples

after drying. This, and the automation offered by every Genevac evaporator system, helps users to deliver

uniform results every time - increasing productivity in environmental analysis.

A series of technical papers demonstrating the advantages and benefits that SampleGenie technology

offers to environmental laboratories

may be downloaded from

www.genevac.com/CFA.

Genevac, part of the SP Scientific

group, was founded in 1990. Today,

the company employs

approximately 85 people, with

manufacturing, R&D and marketing

headquartered in Ipswich, UK.

Genevac offers a comprehensive

portfolio of evaporators to suit

almost any solvent removal

application, purchasing budget or

productivity requirement.

For further information please

contact Genevac on +44 (0)1473-

240000 / +1-845-255-5000 or

email salesinfo@genevac.co.uk

7 PharmaNEWS


ISSUE 1 2010

The Society for Biomolecular Sciences and the Association

for Laboratory Automation announce proposed merger

8 PharmaNEWS

Members to vote on uniting as sections of the new

Society for Laboratory Automation and Screening

For the past 18 months, the Society for

Biomolecular Sciences (SBS) and the Association

for Laboratory Automation (ALA) have been

strategising a merger to unite their complementary

missions within the context of one inclusive

organisation - the Society for Laboratory

Automation and Screening (SLAS). The SBS-ALA

merger awaits membership approval via a proxy

voting process. If approved, SLAS will become the

premier international community dedicated to

advancing scientific research and discovery through

laboratory technology.

“SBS and ALA are facing one of the most

important steps forward in the evolution of our

organisations. Despite serving uniquely different

scientific niches, our members have very similar

educational interests and member service priorities.

The SLAS merger will capitalise on both to provide

increased opportunities for success,” says Jeff

Paslay, Ph.D., SBS President. “We highly encourage

our members to vote yes

for the merger.”

With member approval, SBS and ALA will unite

as individual sections of SLAS, each retaining its

individual identity and specialised educational

focus. While they will also retain their individual

missions as sections, they will collectively address

the overall SLAS mission - to provide forums for

education and information exchange to encourage

the study of and improve the science and practice of

laboratory automation and screening. In addition,

both SBS and ALA will benefit from the expanded

scope, international influence and enhanced

program and service offerings that the unified

umbrella organisation, SLAS, will be able to

provide.

“Our organisations have such complementary

missions that coming together on a broader level

will expand our global reach and influence,” says

Malcolm Crook, Ph.D., ALA President. “Through

the alliance we can leverage each other’s strengths

to achieve international significance and attract new

members who offer increased diversity of

knowledge and experience.”

Voting opens on 15 March at www.SBS-

ALAmerger.org and closes on 5 May. Upon SBS

and ALA member approval, the merger will become

official 1 July 2010. For more information on the

SBS-ALA merger and how to vote, visit www.SBS-

ALAmerger.org

About SBS

Headquartered in Danbury, CT, the Society for

Biomolecular Sciences (SBS) is the only

international non-profit scientific society dedicated

to drug discovery and its related disciplines. SBS

was founded in 1994 to provide a forum for global

education and information exchange among

professionals in the chemical, pharmaceutical,

biotech and agrochemical industries. SBS members

represent many of the largest and most influential

research institutes, universities and pharmaceutical

companies in the world.

About ALA

The Association for Laboratory Automation (ALA)

is a worldwide organisation representing leaders in

all aspects of laboratory automation. ALA seeks to

provide a greater understanding of the importance

and value of automation strategies and technologies

in the laboratory settings, to advance science and

promote education related to laboratory automation.

About SLAS

The Society for Laboratory Automation and

Screening (SLAS) is a new non-profit scientific

organisation uniting the Society for Biomolecular

Sciences (SBS) and the Association for Laboratory

Automation (ALA). The SLAS mission is to be the

preeminent global organisation providing forums for

education and information exchange to encourage

the study of, and improve the science and practice

of, laboratory automation and screening.

www.europeanpharmaceuticalreview.com


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Dr Sheraz Gul

Vice President and Head of Biology,

European ScreeningPort, Hamburg

Article 1, 2010: Lab

Automation series

In this first article of 2010 for the Drug Discovery Developments section of European Pharmaceutical Review

Digital, an overview of the common types of assays as well as emerging trends for the protease enzyme target

class, which are responsible for the catalytic cleavage of peptide bonds in their substrates, will be covered. The

dysfunction of many of these enzymes is known to be the cause of a number of diseases and a number of

protease inhibitors have been approved for clinical use. With the advances in assay methodologies that have

recently been made, protease assays that make use of physiological substrates rather than peptide substrates

(whether physiological substrate derived or generic) can now be readily employed in compound screening

activities.

9 Lab Automation

The protease enzymes are a well

validated target class in drug

discovery, with many drugs

having been approved for

clinical use whilst many other

proteases are currently being

investigated with a view to

discover novel compounds that

modulate their catalytic activity

and can be potentially used as

drugs. There are a number of

types of protease enzyme and

these are generally classified

depending upon the nature of

the catalytic machinery within

the catalytic site that is

responsible for catalysis. Thus,

the most common members are

the serine, cysteine, aspartyl and

metallo proteases. As proteases

are found ubiquitously in living

organisms, often in significant

amounts in plants in particular,

proteases from these sources

have historically been studied in

academic research groups for a

long time. Although the plant

proteases are not directly of

therapeutic value, studies

making use of them have

provided extremely valuable

insights into protease

mechanism of action, specificity

characteristics, protein folding,

structural information and the

design and development of

substrates and modulators of

their activity. The principles

arrived at from the study of such

model proteases have had a

direct and beneficial influence

on those proteases of

therapeutic value.

A number of strategies can

be explored when protease

assays need to be designed

and subsequently employed in

compound screening activities.

In the case of the protease

enzyme itself, it is desirable to

make use of full length protein

that has been expressed in a

suitable system so as to

generate the protein in a form

that is as physiologically

relevant as possible. A tag is

also often expressed at the


ISSUE 1 2010

10 Lab Automation

www.hecus.at

termini of the protein to facilitate its

purification and this should be removed by

the inclusion of a suitable cleavage site

between protein and tag. In the case of the

substrate for the protease enzyme, a number

of options are available and careful thought

needs to be given as to which of the possible

substrates should be utilised for assay

development and subsequent compound

screening activities. The importance of

choosing the most appropriate substrate

stems from the fact that protease enzymes

contain an active centre which is composed

of a catalytic site where the residues directly

involved in the cleavage of the substrate are

located and a binding site where the

substrate can interact with the protease

enzyme. It is usually the binding site that

confers substrate specificity characteristics

upon the enzyme. In many proteases, the

binding site accommodates multiple

residues of the substrate on either side of the

scissile peptide bond of the physiological

substrate. Early studies on proteases made

use of simple chromogenic substrates such

as p-nitrophenylacetate. Although this

substrate allows the detection of protease

activity, it contains no specificity features for

any particular protease. This is a problem as

the measured activity may be due to other

contaminant proteases within the

preparation and if the assay is used without

sufficient validation in compound screening

activities to identify starting points for drug

discovery, the use of such an inappropriate

www.europeanpharmaceuticalreview.com


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substrate may lead to the

allows them to be used

nm. Alternative fluorogenic

identification of modulators of

without any separation steps

substrates make use of

protease activity whose

and in most cases can be used

quenched peptides that

properties may not translate to

to monitor protease activity

contain a donor and quencher

more physiologically relevant

kinetically and in real time.

chemically coupled to residues

assays which the compound

must be shown to be active in

for drug discovery projects. The

use of peptide based

substrates that have more of a

resemblance to physiological

substrates therefore offers

significant advantages over

p-nitrophenylacetate as they

Chromogenic substrates

contain a group which when

released after the protease

mediated cleavage gives

rise to colour change

which can be followed

spectrophotometrically and the

signal is proportional to the

protease enzyme activity.

within the peptide substrate

which are in close spatial

proximity. An example of such

a substrate is one that contains

the fluorescent AMC group that

is quenched by resonance

energy transfer to the 2,4-

dinitrophenyl (DNP) group.

Upon catalytic cleavage of the

11 Lab Automation

will incorporate some of the

Chromogenic protease

peptide, these two groups are

specificity requirements of

substrates are not ideally suited

spatially separated, quenching

the enzyme.

for compound screening

is overcome and the extent of

Early peptide substrate

activities due to their lack of

product that is produced can

based protease assays made

sensitivity. In the case of

be determined by exciting the

use of unlabelled substrates

p-nitrophenyl derived

system at 320 nm and

which, after undergoing

peptide substrates, the

detecting the emission at 405

cleavage to generate lower

chromophore that is produced

nm. In the case of the

molecular mass products,

(p-nitrophenol) is monitored by

chromogenic and fluorogenic

would be separated by

measuring the absorbance at

substrates, the wavelengths

methods such as thin layer

410 nm. The commonly used

involved in the assays are in the

chromatography only after

fluorogenic substrates (which

region where compound

which the activity of the

are significantly more sensitive

interference is likely so care

protease could be calculated.

than chromogenic substrates)

needs to be exercised when

However, such assays are very

contain the 7-amino-4-methyl

interpreting the results from

low throughput and not

coumarin (AMC) group. Upon

such assays. The fluorogenic

compatible with screening

catalytic cleavage of the

peptide based substrates that

large libraries of compounds.

substrate, the product is

are commonly used in

Thus the use of appropriately

detected by excitation of the

compound screening activities

labelled peptide substrates

system at 380 nm and

are an improvement upon

(chromogenic or fluorogenic)

detecting the emission at 460

chromogenic substrates;


ISSUE 1 2010

however, they still have the

enable the development of

such as cell based assays where

unnatural fluorophores within

proximity based assays such as

the activity of the protease can

the substrate molecule that

Time Resolved-Fluorescence

be assessed against the native

could compromise the binding

Resonance Energy Transfer

substrate. It is therefore

modes with the enzyme.

(TR-FRET) or Amplified

essential to develop protease

12 Lab Automation

Although robust assays can be

developed when using

methodologies that employ

fluorophore labelled substrates,

the compounds that are

identified from such assays

may exhibit a profile that does

not translate to a more

physiological assay. The use

of such compromised assays

may in part explain the

Luminescence Proximity

Homogeneous Assay

(AlphaScreenTM) technologies,

both of which are compatible

with the commonly used

micro-titre plate formats.

However, a prerequisite to

developing these types of

assays are the appropriate

labelling of the antibodies.

It is now becoming easier to

assays that make use of

physiological substrates in the

first instance. This shift in

strategy should facilitate

the reduction in attrition in

drug discovery.

Contact the author

sheraz.gul@screeningport.com

high attrition rate associated

set up biochemical assays for

with the early stages of

proteases using reagents that

drug discovery.

can be purchased

As the catalytic action of

commercially. These assays can

proteases upon their substrates

subsequently be employed in

produces products that have a

high throughput compound

newly generated epitope that

screening activities to identify

is absent in the initial substrate

molecule, this feature can be

exploited in the development

of a protease assay. If the

substrate is a protein and the

cleavage site within it is known

and if antibodies are available

that specifically recognise the

newly generated epitope, its

use with another antibody that

recognises the total protein e.g.

N- or C- terminus antibody, will

compounds of potential

interest in drug discovery

efforts. Many of these protease

assays make use of peptide

based substrates that are nonnative

(whether derived from a

physiological substrate or a

generic substrate) and profiles

of compounds identified from

such types of assays all too

often do not translate to more

physiologically relevant assays

Dr Sheraz Gul

Dr Sheraz Gul is Vice President and

Head of Biology at European

ScreeningPort, Hamburg, Germany. He is

responsible for the management and

development of Medium and High

Throughput Screening activities for

academic partners across Europe. He has

12 years research and development

experience in both academia (University

of London) and industry

(GlaxoSmithKline Pharmaceuticals). This

has ranged from the detailed study of

catalysis by biological catalysts

(enzymes and catalytic antibodies) to the

design and development of assays for

High Throughput Screening for the major

biological target classes. He is the

co-author of numerous papers, chapters

and the Enzyme Assays: Essential Data

handbook.

www.europeanpharmaceuticalreview.com

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