Developing Better Assays for Every Human Protein

OriGene Technologies, Inc.

Mark E. WatsonSr. Director Business Development

Phone: (240)994-6873Email: mwatson@origene.com

Collaboration Opportunities with OriGene

• Strategic Licensing Agreements

• Supply agreement for cDNA clones, full-length human proteins and validated monoclonal antibodies

-Purchase of full cDNA clone collection or subsets-Human and Mouse ORF clones available -Bulk full-length protein development -Licensing of hybridoma cell lines (e.g. TrueMAB mAbs)-Joint mAb and/or Hybridoma cell line development -Custom multiplex assay development

• Collaboration agreements for antibody decoding and auto-antibody studies using the proprietary protein lysate microarray technology from OriGene

• Custom service projects leveraging gene synthesis capabilities from Blue Heron Biotechnology, including codon & antibody optimization

OriGene’s Key Capabilities• Largest collection of cDNA clones in the world • Protein production in human cells

– 12,000 transfected antigen lysates– 5,000 purified proteins from HEK293Tcells– Est.10,000 available by the end of 2011– Heavy-labeled MRM/Mass spec. standards.

• High throughput mAb production/ validation – Recently opened second facility to expand production to over 500 released and fully

validated mAbs/month– Three fold increase in production capabilities by year end 2011

• Cytomyx tissue bank: – >140,000 tissue samples – High throughput IHC validation capabilities

• Synthetic Gene Production & Services– Blue Heron Biotechnology Acquisition (Ab & Codon optimization)

• 10,000 Antigen Chip (Autoimmune & Cancer tests)• Luminex multiplex assay development (Marligen acquisition)

Complete Gene/clone Resource

Clones/genes Proteins Antibodies Assays

TrueORF/GoldHuman/mouseExpression vectors

True Proteins5,000 humanCustom purification

TrueMAB>1,800 mAbsIHC, IF, IP, FC, WB

TruePlex

Any speciesAny sequencesAny variants

Optimized expression

Antibody engineering Multiplex Assays

High Throughput Work Flow

15,000TrueORF Clones

12,000Over-expression Lysates

5,000Purified Protein

Transient Transfection(MegaTran)

Cell Lysates AffinityColumn

Largest offering of mammalian cell expressed human proteins

Highest Quality Standard

Not just sequence-verification

But also,

Expression Validation on 12,000 TrueORF clones

APO3C

TrueORF:RC206566

ORF size: 300 bp

Predicted MW: 11 kd

TP53

TrueORF:RC200003

ORF size: 1182 bp

Predicted MW: 53 kd

ENC1

TrueORF:RC201842

ORF size: 1770 bp

Predicted MW: 65 kd

LRP1

TrueORF:RC218369

ORF size: 13635 bp

Predicted MW: 504 kd

CDH3

TrueORF:RC207346

ORF size: 2490 bp

Predicted MW: 91 kd

Transfected Over-Expression Lysates

• 12,000 TrueORF clone transfected HEK293T cell lysates

• Mammalian cell expressed full length, WB validated

• Delivered in modified RIPA buffer to preserve protein function

• Ideal tools for antibody validation, protein function studies, and as assay standards

Over-expression Lysates

• OriGene over-expression lysates adopted in Antibody Atlas Program to validate polyclonal antibodies (e.g. Atlas Antibodies).

• Improved validation success rate from 32% with regular cell lysates to 89% with over-expression lysates.

• Data from Prof Mathias Uhlén, Program Director.

Using OriGene antigen microarray chip technology to decode antibodies that were generated by whole cell immunization

Anti-DDK (Rabbit) Anti-E-Cadherin1 (mAb)

Enlarged image

E-Cadherin1

10,000 Lysate Protein Arrays

Full Length Human Proteins

• Produced from TrueORF cDNA clones

• 5,000 full length human proteins

• Expressed in HEK293 cells

• Optimal preservation of protein structure, post-translational modifications and functions

Human Protein Activity

mTOR Activity Assay

Homogeneous time-resolved fluorescent (HTRF®) assay.

mTOR is a serine/threonine protein kinase that regulates cell growth, cell proliferation, cell motility, cell survival, protein synthesis, and transcription. mTOR belongs to the phosphatidylinositol 3-kinase-related kinase protein family.

First commercial provider of active, full-length human mTOR protein

MS Collaboration with ISB

• Generate signature peptide sequences for all 5,000 human proteins – Sequences will be in PeptideAtlas database

• Heavy-isotope labeled purified proteins– 5,000 available now for SRM/MRM application

• Isotope labeled peptides –Coming in 2011 for all 5,000 human protein

mAbs for Every Human Protein

Dedicated mAb production center - Capacity: +200 immunizations per month

Authentic proteins as antigens- Full length proteins made in human cells

Extensive validation- WB: 9 different cell lines- IHC: 24 different tissues - IF- Flow cytometry- Luminex- Additional validation available

Hybridoma cell lines for licensing- High value targets available: ERCC1, LGR5,

HES1 & NOTCH1

Goal: To produce high quality mAbs to every human proteins in 5-10years

HTP Validation of mAbs

IHC data for human carcinoma liver tissue

Anti-KRT WB with OriGene over expression lysate

Anti-KRT WB with cell panel of 9 different cell lines(For endogenous protein detection in cell line)

ERCC1 (4F9)ERCC1 (1A3)IF data with A549 cells IF data with A549 cells

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Analyte Concentration (pg/ml)

FGF -4

VEGF -D

EGF

HGF

PlGF

VEGF -A

Med

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Quantitation of 6 Growth Factors in an 11-plex ELISA

Immunoassay Development

Summary

OriGene has developed the most comprehensive genome wide product solutions available

Leading developer of high quality reagents and multiplex assays for every human protein

OriGene is your perfect partner for companion diagnostic and biomarker assay development

Additional information available at:

www.origene.com