1. ProteoGenix Life Sciences Services and Products
From gene to biotherapeutics
Target Validation to Lead optimisation

2. ProteoGenix Philippe FUNFROCK, founder and CEO
French company located in Strasbourg, 14 years of existence
One stop solution from Gene to Diagnostics / Biotherapeutics, Food enzymes, or Veterinary products…

3. Antibody drug market
Major class of biopharmaceutical products worldwide
First in 1986 (Muromonab-CD3)
USD 85.4 billion in 2015
More than 500 in the clinic
murine IgG2a CD3 specific transplant rejection drug, OKT3 (also called muromonab), in 1986

5. Context Antibody drug discovery  sophisticated process
Time to the market increase
Significantly more costly to develop and produce
Begin with the End in Mind. Stephen R. Covey

6. Context
The better the hit antibody initially meets the specifications  the higher the probability to make it to the market
These antibody properties depend on how the antibody is developed

7. Specifications of a therapeutic ab
Format (100+)
Functionality (activating, inhibiting, blocking)
Specificity
Species cross-reactivity (mouse, monkey)
Stability / solubility
Affinity
Productivity
ADA response
2015 :
Antibody domain structure has been exploited to make >60 alternative BsAb formats.
2 BsAb are approved for therapy and >30 BsAb are in clinical development.
•A common clinical application of BsAb is retargeting T cells to kill tumor cells.
•Other clinical uses of BsAb include dual blockade of different disease mediators.

8. Formats Full-length mAbs Bi-specific mAbs ADC sdAb (VHH)
Other (ScFv, Fab, polyclonal, Fc-fusion proteins…)
omab (murine, no more used)
ximab (60% human)
Zumab (90% human)
Umab (100% human)
Bi-spe in 2015:
ADC in 2015:
superior pharmacological efficiency along with minimized side effects
By attaching biologically active chemotherapeutic drugs, radioactive isotopes, cytokines or cytotoxins via chemical linkers with liable bonds to a monoclonal antibody
about 50 molecules are in clinical development

9. 100+ BsAb Formats
International experts

10. How to develop therapeutic mAbs FlowChart example

11. Antigen (Target) choice
The choice of the right antigen is decisive
Functionality, affinity and specificity
Native Protein, Recombinant protein, peptide, DNA (Genetic immunization)
Endogenous protein highest similarity
Full-length, fragment…

12. Gene synthesis To produce the antigen (target)
Define the expression system
cDNA optimization
Vector choice
Promoter, leader sequence
Full-length vs fragment
Domain to produce, SIP, tag position, cleavage site
Leader sequence :
Avant 5’. 5’ Uter ans translated region + kozak.
Kozak helps for transcription (transform gene into mRNA)
Other help for traduction (mRNA to protein): ribosome binding site, hairpin…

13. Antigen (target) production
Cell line (HEK293, CHO)
Tricky proteins  High throughput
SIP, tags, medium, culture conditions, feeding

14. Antibody development (hybridoma)
Large number of clones required
Mice and fusions. Higher probability of having the expected features
Testing at parental stage
Subclone best clones
Screening in several applications
Hybridoma sequencing

15. Antibody development (hybridoma)
Produce the recombinant version in HEK293, CHO cells or E.coli
Compare affinity and functionality of hybridoma and chimeric recombinant version

16. Antibody library screening Phage display
Pros: Selection of Human/VHH and recombinant, time, efficiency, toxic molecules
Cons: affinity
Naïve or Immune library screening (Fab, ScFv, VHH, species…)
Key factor: quality (diversity) of the library

17. Antibody engineering Humanization Format development
CDR grafting, reduce ADA
International experts
Format development
Stability improvements
3D modeling

18. Affinity Maturation Random mutagenesis Phage display
Diversity up to 10^8 of new library
Phage display

19. Hits characterization
ELISA, WB, FACS
Cross-reactivity to mouse and monkey targets
Generate surrogate antibodies
Functionality analysis (in vitro)
Aggregation
Functionality in cell-based assay
In drug development, Pharmacokinetic (PK) analysis of the drug is performed to determine the fate of the drug (e.g., distribution, absorption, and/or secretion) after administration to an animal or human. This analysis is often carried in conjunction with Pharmacodynamics (PD), the study of a drug’s pharmacological effect on the body. The PK and PD assays typically require critical reagents (e.g., specific anti-drug Abs) and the development of an assay that can measure the drug in plasma and tissues.

20. Leads characterization
Affinity determination
Biacore, OctetRed, SPRi
Epitope mapping
ADCC, CDC
PK/PD analysis
In vivo efficacy
Toxicity studies

21. Stable cell line development
Vector technology and transfection
Choice of the cell line (CHO)
IP conditions
Selection marker (GS, DHFR)
Productivity several g/L
GMP transferable

22. Process development Bioreactor production
Medium optimization
Composition, anti-foam and shearing aspects
Feeding strategies
Process monitoring
pH, T°, gas streams, side components, product…
Purification optimization
Increase recovery rate, quality and purity
Fine-tuning
Side components: resulting metabolites, wastes
Product: expression level, plateau…
Fine-tuning: EU, DNA, Host cells removal, formulation, sterilization, lyophilisation

23. Our services Protein production : 1 000+ pAbs production : 1 500+
5 expression systems, stable cell line, fermentation, process development, High Throughput screening...
pAbs production :
Peptide synthesis :
Gene synthesis :
Starting 0,21€/bp, Gene optimization tool
mAbs development : 200+
Hybridoma technology, Phage display, Antibody Engineering (Humanization, Bi-specific, ADC, ScFv, Fab, Fc-fused protein)

25. Why choose ProteoGenix ?
One stop solution: Multi-services association experience l
increased success rate
More efficient troubleshooting
Reduced lead time
Speed
Strong team technical background and experience
High levels of Guarantee
 Visit our booth

26. Thank You Your contacts: ProteoGenix Philippe Funfrock Founder and CEO
Emeline Ludwig
Business Developer
ProteoGenix
15 rue de La Haye
67300 Schiltigheim
France
Phone: