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One-Stop Antibody Drug Discovery Services from GenScript ProBio PowerPoint Presentation

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Slide 1 - Inspire, Accelerate & Co-create Biomedical Innovation
Slide 2 - 1 www.GenScriptProBio.com GenScript ProBio GenScript ProBio is the bio-pharmaceutical CDMO segment of the world's leading biotech company GenScript Biotech Corporation (Stock Code: 1548.HK). Founded in 2002 in New Jersey, GenScript started business from gene synthesis. Now, GenScipt ’s businesses encompass four major categories based on its leading gene synthesis technology, including operation as a Life Science CRO, enzyme and synthetic biology products, biologics development and manufacturing, as well as cell therapy. In Jan 2019, GenScript established the Biologics Development Business Unit (BDBU) which is the predecessor of GenScript ProBio. The name of "ProBio" indicates 3 core philosophies - being PROACTIVE, PROFESSIONAL and PROCESS-oriented. GenScript ProBio shows our dedication to proactively provide end to end service (discovery to commercialization) with professional solutions, and efficient process to accelerate drug development for customers. GenScript ProBio provides an integrated biologics discovery & development solution from target to IND. With our cutting-edge technology platforms in therapeutic antibody discovery & development, GenScript is able to deliver functional antibody lead with good developability and safety in discovery phase, as well as reliable, productive and regulatory-compliant process & drug product for IND filing in development phase, which significantly save client's time and cost. Antibody humanization Affinity maturation Developability assessment Bioassay & Bioanalytics Lead Generation Hybridoma generation Human naïve library & Synthetic library Fully human transgenic mice Single B cell cloning SMAB bispecific antibody discovery Lead Optimization Biologics Development Cell line development Process development Analytical development Clinical batch supply
Slide 3 - www.GenScriptProBio.com 2 GMP Plant #1 Nanjing, China GMP Plant #2 Nanjing, China GMP Plant #3 Zhenjiang, China Leading Therapeutic Antibody CDMO From Target to Market Solutions 500+ Employees, 40+ Experts with extensive experiences in industry cGMP compliant Mammalian Cell Culture Facility Comprehensive Analytics Platform
Slide 4 - 3 www.GenScriptProBio.com GenScript's 13 years of custom antibody generation experience can deliver a panel of hybridomas under 4 months with our standard protocol. We Provide A full spectrum of immunization approaches: Protein, peptide, whole cells and DNA. OptimumAntigen™ Design Tool guarantee results. Optimized immunization:Transgenic mice* generate human antibodies. Proprietary adjuvant and immunogen modification breaks immunological tolerance for superior immune response. Complete service: From antigen production to hybridoma development and characterization. High throughput: High efficiency screening. Proprietary NativeSelect™ ELISA for soluble targets. iQue HT Screener, and BD FACS Calibur with HT loader for membrane targets. Comprehensive functional assays: Validated functional assay platforms provide reliable in vitro screening. Readily integrated with other drug discovery services: Antibody sequencing, antibody humanization, recombinant antibody production and anti-idiotype antibody generation. Note: We can customize your project based on your specific requirement. We provide free cell line storage service for 6 months from the date of cell freezing. We recommend GenScript provides immunogen for highest quality . * Hybridoma Antibody Drug Discovery Target Animal immunization Customer function assay Customer function assay 1-5 mg mAb/clones 50-100 clones 100 clones 40000 clones Test bleed report Top clones for antibody engineering Binder clones subcloning & production SuperdomaTM assay & lsotyping High throughput ELISA & FACS screening Cell fusion
Slide 5 - Human antibody sequence in 2 months Fully human antibodies are the mainstream of therapeutic antibody development because of the increasing concern on immunogenicity and safety issues. Among the techniques that develop fully human antibodies, Human Naïve Library and Phage Display have been proved* to be an effective platform to pave the way to the fully human candidates. GenScript has successfully developed in-house Human Naïve and Llama Naïve Library, which have been internally validated for multiple targets. Besides, our experienced scientists will develop multiple panning strategies, which are just right for you to get the best antibody candidate. 4 Technical Indicator Human Naïve Library Llama Naïve Library for sdAb (Available Now) Source Library format Library size Insert rate In frame rate/ORF rate CDR3 diversity Germline analysis 2400 healthy donors Fab-HK Fab-HL 1.04*1011 (In Expanding) >90% >85% Normal distribution Nature frequency 100 healthy alpaca donors sdAb 2*1010 (In Expanding) / >95% Normal Distribution Nature Frequency Phage Display and Antibody Library Advantages Liquid panning Solid panning Cell panning Ag.biotin-cell panning Ag-cell panning Competitive panning Better at presenting all possible epitopes, high diversity Simple format, suitable for most target Generating antibodies able to recognize natural epitope presented by cells Generating antibodies able to recognize more diversified epitopes and natural epitope presented by cells Generating antibodies able to recognize natural epitopes presented by cells Eliminate antibodies targeting irrelevant epitopes, acquiring antibodies specific to desired epitope GenScript’s Naïve Library and Phage Display Service: Panning: Diversified panning methods to increase the successful rate. Panning strategies tailored to different targets: soluble and transmembrane targets. No more than 3-round panning to retain the sequence diversity. Screening: A combination of FASEBA high throughput platform, ELISA/FACS screening, and SPR affinity screening. *Adalimumab, Belimumab, Raxibacumab, Ramucirumab, Necitumumab, Avelumab, Ranibizumab and Durvalumab are known to be developed by phage display antibody library technology. www.GenScriptProBio.com
Slide 6 - 5 www.GenScriptProBio.com Single B Cell Screening for Antibody Drug Discovery Antigen ligand (positive control antibody) binding/blocking with ELISA and/or FACS, EC50 measurement, optimize antigen concentration and/or secondary antibody concentration Option 1: Immunization with protein (regular animals) Option 2: Immunization with protein (Transgenic animals*) Protein-based IgG Protein-based antigen specific binding Cell based assays(epitope mapping, antigen binding, ligand blocking, etc) Desirable single B cells (given the selection criteria) will be exported for total cDNA recovery and single cell VH/VL sequencing Ab expression & purification 1-2 Weeks 1 Week 1 Day Assay on Single B System: Beacon Sequencing Recombinant Antibody Production Pilot Experiment 8-14 Weeks(Standard) 2 Weeks(Express) EXPEDITED TIMELINE Screening completed in 1 day As fast as 1 month to get functional sequences INTRINSIC ADVANTAGE for CHALENGING TARGET High hit rate: multiple screening/assay method to select functional leads at earlier stage Low hit rate: limited B cell repertoire due to fusion loss MAXIMIZED DIVERSITY Minimal diversity loss, forward functional screening HUMAN SAMPLE COMPATIBLE Full human antibody screened directly from human PBMC Suitable for infectious disease research Beacon IMPORT EXPORT MULTIPLEX ASSAY Antibody Secreting Cell Enrichment B Cell Harvest Immunization Antibody Expression And Purification Single B Cell Sequencing 1 day GenScript ProSpeedTM Service: suitable for soluble targets, single transmembrane targets and also multiple transmembrane targets. Human PBMC samples could also be used as starting material. Phase Item TAT Materials Preparation Antigen preparation Finish Antibody Screening within 24 hours Single B cell screening platform, also known as single B cell cloning technology, is a microsystem based screening method. This platform conducts isolation, screening and evaluation on the B cells, avoiding the cell fusion and the library construction step. Also, single B cell screening platforms usually integrate high throughput platforms, therefore assays could be carried out in a highly efficient manner. GenScript has built our own single B cell platform by using Beacon™, a micro-chamber based platform. With dozens years of experience on antibody discovery and cell manipulation, GenScript now provides a one-stop solution on antibody discovery using single B cell screening technology. GenScript ProSpeedTM Service Features *
Slide 7 - www.GenScriptProBio.com 6 Antibody Sequencing Our Experienced Hands, Your Unique Sequence GenScript takes the greatest care to ensure a 100% accurate sequence. Each chain is cross verified amongst 5 independent clones. No mutation is introduced with both 5’ and 3’-RACE Full Length sequencing of the variable region from FR1. Value and Speed Within duration as short as 6 days, GenScript delivers the data needed with your intellectual property being protected. Sequencing of antibody binding regions also sheds light upon further development such as humanization and recombinant antibody production. Sequencing of Variable Domain/Full Length IgG Clone into Standard Sequencing Vector Optional: Clone into Expression Vector Optional: Recombinant Antibody Expression Optional: Antibody Binding Confirmation Starting Material from Clients: >1×106 cells
Slide 8 - Reduce the Immunogenicity, Keep the Accuracy Replacing everything but the complementarity determining region (CDR) reduces the degree to which an antibody drug itself acts as an immunogen. Immunization against an antibody drug lowers efficacy through reduction in circulating half-life and/or neutralization. Our Strategy CDR-grafting Patented library based “framework assembly” FASEBA: FAst Screening for Expression level, Biophysical properties, and Affinities Case Study Turnaround Service Milestone Deliverables Chimeric antibody production and binding confirmation Antibody Humanization The engineering and screening procedures are integrated, allowing for simultaneous antibody humanization and stabilization. We even guarantee the affinity of the humanized antibody, which will be equal to or higher than the affinity of your parental antibody. 0 200 400 600 0 20 10 30 Response(RU) -4 -8 -12 0 4 8 217nm 208nm 202nm CD (mdeg) 42 52 62 72 Temperature (℃) 82 92 -4 -8 -12 -16 4 0 8 217 nm 208 nm 202 nm CD (mdeg) 42 52 62 72 Temperature (℃) 82 92 nM nM 4 nM 8 nM 16 nM 2 nM Repetition nM nM 4 nM 8 nM 16 nM 2 nM Repetition D -10 K = 7.22×10 M Mouse IgG Humanized IgG D -10 K = 4.92×10 M Time (s) Tm = 70.6 ℃ 600 0 0 20 10 30 200 400 Time (s) Tm = 66.3 ℃ Response(RU) Express Humanization Sequence, DNA and chimeric Ab & top 3 humanized Abs (0.5 mg) Guarantee at least 1 Ab with affinity comparable with parental Abs Sequence, DNA and chimeric Abs & top 3 humanized Abs (0.5 mg) Guarantee at least 1 Abs with affinity comparable with parental Abs 8 Weeks 11 Weeks Express humanization by rational design and sequence synthesis Humanized antibody production and affinity ranking Humanized antibody production and affinity determination Chimeric antibody production and Binding confirmation Humanized antibody production and characterization 7 www.GenScriptProBio.com Sequence PTM analysis, Back mutation library design and library construction Humanized antibody selection PTM analysis（optional） Deluxe Humanization
Slide 9 - www.GenScriptProBio.com 8 5-100 fold of Affinity Maturation Affinity is one of the key parameters of an antibody drug, which will affect the function and efficacy of the antibody. Generally, antibody candidates from hybridoma platform has already acquired high affinity, but it may not exactly fit in the practical needs in research. GenScript provides Affinity Maturation Service, which will help to improve the antibody affinity as you desired. Affinity Maturation Case Study Affinity maturation was performed for the Wild Type IgG and 3 positive clones were found, with Clone 1 showing highest fold increase(751 fold increase). Paratope Mapping Single Point Mutation Library Design and HTP Screening Affinity Matured IgG Production and Characterization Core Advantages Guarantee the 5-10 fold increase Application of FASEBA High Throughput platform, in which affinity, expression level and thermal stability are screened simultaneously. Service Details Deliverables Timeline Antibody sequencing and sequence analysis Paratope mapping, library design and HTP screening Antibody and sequence and report analysis Top 5 clones Guarantee 5-10 fold affinity improvement 20-24 weeks
Slide 10 - 9 www.GenScriptProBio.com Seamless Connection from Antibody Discovery to Preclinical CMC Development Some antibody candidates discovered in early stage will involve issues such as post translational modification (PTM) hotspots or poor physiochemical stability, which will lead to potential risk during preclinical CMC development, resulting in huge time and financial loss to researchers. Taking advantage of bioinformatics tools and a series of quality study instruments, GenScript introduces Developability Assessment Service to identify the inherent risk in antibody discovery stage, which seamlessly connects antibody discovery with preclinical CMC development. Preliminary Developability Assessment Chemical Stability Physical Stability Developability PTM Analysis Fragmentation Deamidation Isomerization N-Glycosylation Oxidation Free-thiol PTM Hotspots Service Package Service Content Timeline Physiochemical assessment PTM hotspot identification & validation Thermostability Aggregation Hydrophobicity Molecular weight (de-glycosylated and glycosylated) Stability (Freeze-thaw cycle, low pH, 40℃ accelerated degradation, matrix effect) Asparagine deamidation Aspartate isomerization Tryptophan oxidation Hydrolysis N-glycosylation 8 weeks 8-10 weeks
Slide 11 - www.GenScriptProBio.com 10 Why Single Domain Antibody? Potential ability to bind “hidden” epitopes GPCR, ion channel, etc. High affinity: can reach pM range GenScript Proprietary Bispecific Antibody Platform——SMAB SMAB—Bispecific Antibody Platform Bispecific antibodies (bsAb) have become a focus of interest for therapeutic applications with nearly 85 commercial candidates entering the clinical trials and 3 having been approved to market. GenScript SMAB (Single-domain antibody fused to monoclonal Ab) platform naturally combines the single domain antibody and the monoclonal antibody to make a bispecific antibody in symmetric format. With the design concept of “Keeping Natural”, SMAB platform gives good developability and biosuperiority which are comparable to monoclonal antibody. Single-domain antibody (sdAb, VHH) Proprietary linker Monoclonal antibody (mAb) SMAB (Single-domain antibody fused to monoclonal Ab) + VH H C 1 CL VL CH2 CH3 Favorable biophysical properties Flexibility in modality design Bivalent, trivalent, tetravalent Core Advantages of SMAB Our clients include *: Flexibility to modulate dual target effect to achieve best potency Possibility on ADCC/CDC enhancement Uniqueness of sdAb to target to “hidden” epitope Unique Molecular Flexibility CLD titer > 2 g/L Yield > 60% One step protein A purity > 95% Solubility > 25 mg/ml Stability > 95% after 5 times freeze-thaw PK half life 2-3 weeks Low immunogenicity risk Exceptional Developability 3-5 months for SMAB molecule development 10 months for preclinical development 1 plasmid system for CLD No additional purification No post-production process Efficient Development *Disclosed partners.
Slide 12 - 11 www.GenScriptProBio.com Reduce cell growth rate 1 2 9 10 11 γ β α Ca2+ cAMP Comprehensive in vitro Cell-based Bioassay Platforms Antibody drugs exert their therapeutic effect through different mechanisms of action (MOAs) in human body. Validation of these MOAs in vitro during early discovery is prerequisite to further drug development. GenScript has developed various in vitro cell-based bioassay platforms to accommodate such needs in both academia and industry. Assay methods are readily available for more than 30 hot immune-oncology targets and 9 Fc-γ-Receptors. IND-filing standard can be fulfilled upon request. Cell-based Bioassay Natural killer cell Tissue macrophage Tumor cell GPCR agonist assay Agonist Assay of Epinephrine on ADRA1B Antagonist Assay of Sample A on ADORA2A 120 Growth Inhibition Assay of Sample B on Calu-1 cells Neutralization Assay of anti-IL-6 antibody on STAT3 phosphorylation in TF-1 cells PD-1/PD-L1 Blockade Assay of Anti-PD-1 antibody 4-1BB Agonist Assay of Anti-4-1BB antibody -7 -6 -5 -4 -3 [Anti IL-6 antibody] (mg/ml) (M) EC50 1.109e-005 Neutralization assay Immune checkpoint blockade reporter assay 120 100 80 60 40 20 8000 6000 4000 2000 0 40000 30000 20000 10000 0 800000 600000 400000 200000 0 -4 -2 0 2 % Stim ulation HTRF 665/620 Ratio Lum inescence (RLU) Lum inescence (RLU) % Inhibition % Inhibition 0 10-12 100 80 60 40 20 0 10-12 10-11 10-10 10-9 10-8 10-7 10-4 [Sample A] (M) IC50 3.105e-009 GPCR antagonist assay 100 80 60 40 20 -20 0 -8 -2 10-4 10-4 10-3 10-2 10-1 100 101 102 103 Log10[Anti -4-BB antibody] (nM) EC50 0.7164 Immune checkpoint agonist reporter assay 10-5 10-4 10-3 10-2 10-1 10-0 101 [Sample B] (M) IC50 0.001312 Growth inhibition assay 10-10 10-8 10-6 [Epinephrine] (M) EC50 2.993e-008 Log10[Anti PD-1 antibody] (μg/ml) (M) EC50 0.1014 In vitro
Slide 13 - www.GenScriptProBio.com 12 Peptide 3 4 6 7 8 5 Luciferase RE Growth inhibition assay GPCR-targeted agonist/antagonist assay Neutralization assay Immune checkpoint-targeted reporter gene assay Customized functional assay cell line construction Immune cell stimulation and multiplex cytokine release assay Mixed lymphocyte reaction Complement-dependent cytotoxicity assay Antibody-dependent cell cytotoxicity assay Antibody-dependent cellular phagocytosis assay Cellular characterization assay by flow cytometry T cell Dendritic cell CDC assay MLR assay IFN-γ release Cellular phagocytosis assay MLR assay IL-2 release PBMC stimulation assay ADCC assay IF N-γ (pg/ml) % Target Cell Lysis % Phagocytosis IFN-γ (pg/ml) IL-2 (pg/ml) % Target cell Lysis IFN-γ Release in MLR Assay of anti-PD-1 antibody CDC Assay of anti-Claudin 18.2 antibody Cellular Phagocytosis Assay of anti-CD47 antibody against HL-60 cells IL-2 Release in MLR Assay of anti-PD-1 antibody ADCC Assay of anti-Claudin 18.2 antibody 5000 100 80 60 40 20 0 80 60 40 20 0 2 -4 -3 -2 -1 0 1 [Anti-CD47 antibody] (μg/ml) EC50 0.06014 -20 [Anti-Claudin 18.2 antibody] (μg/ml) EC50 0.3778 3000 80 60 40 20 0 -20 2000 1000 4000 3000 2000 1000 0 10-5 10-4 10-3 10-2 10-1 100 101 102 101 102 10-5 10-4 10-3 10-2 10-1 100 [Anti-PD-1 antibody] (μg/ml) EC50 0.05280 10-5 10-4 10-3 10-2 10-1 100 101 102 [Anti-PD-1 antibody] (μg/ml) EC50 0.009141 [Anti-Claudin 18.2 antibody] (μg/ml) EC50 0.02690 IFN-γ Release from PBMC stimulation assay of anti-Siglec-15 antibody 2000 10-3 10-2 10-1 100 101 102 1500 1000 500 0 10-1 100 101 102 103 [Anti-Siglec-15 antibody] (μg/ml) EC50 38.66 In vitro Cell-based Bioassay
Slide 14 - Note: #Other type: including but not limited to peptide, protein, ADC, SvFv & bispecific antibody. 13 www.GenScriptProBio.com Anti-Idiotype Antibody Generation & Immunoassay Development One Stop Solution from Anti-idiotype Antibody to Immunoassay Development An anti-idiotype antibody (anti-ID Ab) binds to the idiotype (specific combination of idiotopes present within an antibodies complement determining regions) of another antibody, usually an antibody drug. As anti-id Abs specifically bind to therapeutic antibodies, they act as a powerful tool for Pharmacokinetic analysis of antibodies. Anti-ID Abs are also commonly used as a reference standard for anti-drug antibodies (ADAs) in antibody drug immunogenicity (immune response, IG) studies. Features of GenScript Anti-ID Antibodies Service Fast delivery for anti-ID pAb As fast as 8 weeks with guaranteed quantity Application tailored anti-ID mAb Fit for immunoassay development High success rate >98% success rate with > 400 projects Deliverable PK ELISA Kit Development ADA ELISA Kit Development 1-2 mL supernatant/parental clone 5 mL final supernatant/subclone 3-5 Hybridoma cell lines 3-5 purified Ab/clone, 2-5 mg/clone Pilot sensitivity report (optional) COA report Anti-ID pAb Labeled antibody drug Antibody drug 1-2 mL supernatant/parental clone 5 mL final supernatant/subclone 5-10 Hybridoma cell lines 5-10 purified Ab/clone, 2-5 mg/clone, Anti-id mAb pair and pilot sensitivity report (optional) COA report Anti-ID mAb Labeled Anti-Fc Antibody drug Small scale affinity purification (optional) Affinity purified pAb Pilot sensitivity report (optional) COA report Time (weeks) Anti-ID Antibodies Package Details Package Typical Application Starting Material 21-27 21-29 Human IgGI: 8-12 Other type#: >13 ≥ 2 mg target antibody drug 0.5 mg isotype control or human IgG ≥ 2 mg target antibody drug 0.5 mg isotype control or human IgG ≥ 45 mg target antibody drug ≥ 30 mg isotype control or human IgG As detection reagent： Measure antibody drug concentration in patient serum samples As positive control： Measure·ADA concentration in patient serum samples. Measure the level of neutralization antibody, analyze the affect of efficacy ≥ 0.5mg antibody drug ≥1mg detecting antibody ≥ 1mg capture antibody ≥ 1mg antibody drug ≥ 1mg Anti-ID antibody Report of feasibility analysis Final report of kit development Labelled anti-ID antibody Analytical methods and key reagent instructions 8-12 8-12 Labeled Anti-ID mAb Anti-ID mAb Antibody drug Anti-id mAb Golden Package for Capture ELISA Anti-id mAb Deluxe Package for Sandwith ELISA Anti-id pAb Guranteed Package
Slide 15 - Inspire, Accelerate and Co-create Biomedical Innovation
Slide 16 - www.GenScriptProBio.com Email: cdmo@genscript.com Toll-Free: 1-877-436-7274 Tel: 1-732-885-9188 Fax: 1-732-210-0262
 

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