Physiologically Relevant Tools & Personalized Support For Drug Discovery Research

1 10/19/2011 | Life Technologies™ Proprietary and confidential

Physiologically Relevant Tools and Personalized Support for Your Drug Discovery ResearchMark FedericiManager- Strategic AlliancesPSCS-Drug Discovery Assay and Services


Life Technologies brands


Tools for drug discovery & development research

Ste

• Stem cells

• Neurobiology

• Primary cells & matricies

• Cell Therapy Systems (CTS™)

• Target-based proteins and HTS assays

• Kinases• GPCRs• Pathways• Nuclear receptors• Ion channels

• SelectScreen® screening and profiling services

• Custom assay development and cell engineering

• Fresh hepatocytes

• Cryopreserved hepatocytes

•Microsomes

• Transporter products

Cellular Models Target & Lead Discovery ADME/Tox

Our focus today


Technologies & Assays

Screening & Profiling Services

Strategic Alliance Partnerships

• Broad panels of biochemical and cell-based assays

• Primary screening Comprehensive profiling

• Robust Biology Multiple target classes

• Biochemical / Cellular Assays, Primary / Stem Cells

• Customized programs of research (Gene to Screen)

• Dedicated project management

Personalized support for your research

Specialized Technical Support

• Ph.D. level drug discovery focused technical support

• Pre- and post-purchase support


Our team helps our clients develop customized solutions and navigate the full breadth of Life Technologies capabilities

Drug targetprofiling & screening

services

Cell-based assay development

Biochemical assaydevelopment

Biochemical and cell-based ADME assays

Imaging, flow cytometry &cell pathway analysis

Discovery & Target ID

LeadOptimization

SafetyTox

Validation& Lead ID

Preclinical & Clinical Trials

Target-based proteins, assays & services

http://www.invitrogen.com/site/us/en/home/Products-and-Services/Applications/Drug-Discovery/Drug-Metabolism-and-Safety/Biochemical-ADME-Assays.html�


Leveraged Content and Technologies in Professional Services Portfolio

> 1,000 kinase assays –biochemical

> 400 GPCR cellular assays –reporter gene, b-arrestin and second messenger

> 70 Nuclear receptor assays –biochemical and cellular

Multiple Ion Channel technologies and first commercially available Fluorescent hERG assay

> 100 Pathway assays (CellSensor®

and LanthaScreen® GFP)Custom assay development and screening collaborations

Developed the Broadest Target Class Content in the Industry

Integrating products and services

SelectScreen® Services Platform

Library screeningBiochemical & cell-based profiling


Screening, profiling and custom services


From target to preclinical testing: A comprehensive service provider

Targetidentification and validation

Assaydevelopment

Preclinical and clinical

trialsScreening Lead

optimization

Assay Development

Biochemical Assay Development

Cellular Assay Development

Lead Optimization

Molecular and Cellular Profiling

Drug Metabolism CYP450 Profiling

Safety Testing hERG Screening Service

Choose single projects or an entire program

Screening

Bulk Screening Assays/Reagents

Assay Ready Cryo-preserved Cells

Primary and Secondary Screening

Services


Dedicated project management

Single point of contact through dedicated Project Manager with all teams supporting SelectScreen® and Custom Services

R&D

MFG Operations

Dedicated Project Manager

Customer Inquiry

SelectScreen®

Team

MFG Sciences

Custom Biology Team

R&D Custom Assay Team


SelectScreen® platform: A complete solution


Why SelectScreen® services?• Proven track record of rapid, on-time assay development

and screening execution

• Conducted more than 7,600 screening projects on over 400 targets, including library screens with assay development

− Average cycle times:

− 6 days, biochemical profiling

− 8 days, cellular profiling

• Supportive research environment

− Open and frequent communication between the client and Life Technologies dedicated project manager

− Proactive interactions with client based on results obtained

• All of the reagents and assays are developed, manufactured and QC’d on site

• Over 15 years of experience in high quality protein and reagent production, molecular and cellular HTS assay development


Custom service capabilities

Service type Target/functional area

Biochemical assay development Kinases, nuclear receptors, proteases

Cellular assay development Kinases, nuclear receptors, GPCRs, ion channels, epigenetics, pathways

Tb/Eu antibody labeling Kinases, post-translational modifications

BacMam generation Targets and reporters

Stem cells Adaptation, expansion, differentiation

TaqMan® assay development mRNA or proteins

siRNA profiling RNA and functional readouts


Assay platform capabilitiesAssay platform Target area Custom SelectScreen®

service

Z’-LYTE® (FRET) Kinase activity, protease activity √ √

Adapta® (TR-FRET) Kinase activity √ √

LanthaScreen® kinase binding assay (TR-FRET)

Kinase binding √ √

LanthaScreen® biochemical (TR-FRET) Kinase, PTMs, NRs √ √

LanthaScreen® cellular (TR-FRET) Kinase, PTMs √ √

Fluo-4 GPCRs, ion channels √ √

FluxOR™ Ion channels, others √ √

Beta-lactamase/CellSensor™/Tango™ Pathways, GPCRs, NRs √ √

Beta-galactosidase/Luciferase Pathways, GPCRs √ √

TaqMan® assay mRNA √

TaqMan® protein assay Protein: qty, TPM, ix √

Vivid® P450’s and hERG Predictor™ Safety: CYPs, hERG √


Small-molecule vs. siRNA profiling

Feature Small-molecule siRNA

Availability Not available for all targets Synthesize for any target

Mechanism Inhibition of protein function Depletion of protein expression

Timing Rapid, potentially reversible Gradual, sustained duration

Specificity Non-specific effects possible Off-target effects possible

siRNA profiling addresses a broader number of targets and achieves specificity through target-specific knockdown


How can siRNA profiling be used?

1. Dilute oligo

2. Add RNAiMAX

Reverse transfection

ScreeningAssay

3. Add cells

siRNA Library

Gene Expression Analysis TaqMan® assay

Protein ModificationLanthaScreen® cellular assay or

immuno-cytochemistry

Reporter Gene AssayCellSensor® beta-lactamase

Cell proliferationCyQuant® Cell Proliferation assay

or PrestoBlue™


TaqMan® protein assaysProtein Detection from lysates of engineered, normal or primary cells as well as tissues (fresh, frozen or FFPE)

Combines protein detection using Antibodies with robust, sensitive real-time PCR

Relative quantification of proteins in cell and tissue lysates

− No purification of proteins required!

> Just lyse and dilute

Enables very small sample protein analysis

Typical Sample Input

10-500 cells 1-1000 ng protein from tissues

Compatible with StepOnePlusTM, 7500, 7500 Fast, 7900 HT, & ViiATM 7 Real-Time PCR Systems


TaqMan® protein assay features

• Provides all the amplification benefits of nucleic acid based qPCR, but for the analysis of proteins

• Vastly improved sensitivity compared to ELISA and other immunodetection methods, in some cases up to 1000xmore sensitive

• Wide range of exciting new applications:

• Detection of post-translational modifications

• Analysis of both endogenous and exogenous proteins with high sensitivity

• Detection of biomarkers and pluripotency factors

• Detection of protein-protein interactions

• Direct follow-up of mRNA expression with analysis of corresponding proteins levels

p53P

BBB

GFP


Target-based proteins & assays


Comprehensive pathway biology approach

Assay Classes

• GPCR Assays

• Kinase Assays

• Ion Channel Assays

• NR Assays

• Post-translational

modifications

The Integration of Biological Capabilities Into Customer Solutions


Cell engineering tools


What is BacMam?

BacMam uses baculovirus to deliver and

transiently express genes in mammalian cells


How do I use BacMam?

Actin RFP + Tubulin GFP

Clones

TransientTransfections

BacMam

Stable Pools

Workflow

/Assay Tim

e

4-8 Weeks

Fixed Assay

Portable Assay

3-4 Weeks

days

1-2 days

Why should I use BacMam?


BacMam benefits

• Convenient and Fast: just add virus to cells

• Efficient: High expression level

• Flexible: Allows expression of multiple subunits

• Titratable: Control level of expression

• Safe: Non-toxic to cells

• Portable: Works efficiently with most cell types (including primary and stem cells)

BacMam allows you to run the assays you need

in relevant cellular models quickly with high quality results.


Flexibility of BacMam-enabled cellular assays

Bacmam-enabled Cellular Assays enable interrogation of multiple enzymatic activities responsible for posttranslational modifications, including challenging targets like histones.


What is Jump-In?

Jump-In™cell line

Isogenic Jump-In™ Cell Pool

+

AntibioticSelection

R4 Integrase

Gene of Interest

4 weeks

Transfection

For Research Use Only. Not intended for animal or human therapeutic or diagnostic use

Genomic Integration of Genetic Material via Homologous Recombination

The Ideal Solution For:• Building and developing a panel of targets

- All Targets integrated at the same, single genomic site

• Rapid Assessment of Target Biology

Foundational tools:• Jump-In™ platform cell line• Gateway® targetable vector• Integrase vector


Jump-In™ cell engineering technology

Jump-InTM Fast Gateway®

System

Jump-In™ TI™ Gateway®

System

Jump-InTM

Parental Cell Line Kits (New!)

Jump-InTM

Custom Services

Saves time and effort compared to traditional cell engineering methods

Yes Yes Yes Yes

Engineered cell line is isogenic

No Yes Yes Choice

Can use your preferred cell background

Yes Yes Limited to parental cell lines in catalog

Choice

Requires generation of platform/parental cell line (with R4 sites)

No Yes No Choice

Requires Clonal Selection Optional Yes Optional Choice

Ideal when you need… Faster stable cell lines in your preferred cell background

Isogenic cell lines in your preferred cell background

Isogenic cell lines created with less hands-on time

Experts who can meet your project specs and timelines


CHO-K

GripTite™

T-Rex™ CHO

T-Rex™ 293

Jump-In™ cell line portfolio

Currently available through

Discovery Services

HeLa

K562

786-O

HEK293 (2x)

JurkatU2OS

HCT116

TANGO-U2OS

H9

NCI 60

Hemato-poeitic

“Workhorse”

Assay enabled

StemCells

A549

BG01vHEK293

R4

For Research Use Only. Not intended for animal or human therapeutic or diagnostic use

The Jump-InTM cell lines are not catalog products. For more information on how to gain access to this technology, please contact us at [email protected]


Non-excisable integration of construct

Accepts large multigene inserts (up to 20 kb)

Streamlined workflow for cell line generation possible

Stable pools/cell lines easy to maintain

Ideal solution for studying mutants/orthologues:

– Gene of interest expressed from same locus in cells

– Stable pools can be quickly generated

Jump-In™ benefits

Jump-In CCKAR Pool vs. Traditional Clone

-12 -11 -10 -9 -8 -70

500

1000

1500

2000Jump-In 293 CCKAR pool

Traditional CCKAR clone

log10[CCK-8-S] (M)

RFU


Resources & support


Personalized support

Collaboration in action: video portal www.invitrogen.com/collaborate

Drug discovery technical [email protected]

SelectScreen® and custom services [email protected]

http://www.invitrogen.com/collaborate�

mailto:[email protected]�

mailto:[email protected]�


Personalized support

Virtual visits program Sign up at: find.lifetechnologies.com/visits

Free personalized webinars for drug discovery researchers

− Private, 1:1 webinar with Life Technologies R&D experts

− 30 minutes

− Six topics to select from:

> Cellular methods for kinase analysis

> Assay development for primary cells

> Stem cell culture and modification

> Cell engineering

> Assays for drug metabolism

> Research partnerships and services

http://www.lifetechnologies.com/virtualvisits�

Thank you!

Mark [email protected] 908 752 4864

Physiologically Relevant Tools & Personalized Support For Drug Discovery Research

Technology

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