Antibodies by genetic immunisation for therapy and companion diagnostics

Transcription

1 Antibodies by genetic immunisation for therapy and companion diagnostics 5th Munich Biomarker Conference Dec. 1-2, 2015

2 Company overview Contract Service Organisation Aldevron LLC Fargo Established in 1998 Acquired GENOVAC in 2004 Opened Madison facility in 2009 Antibody Technology 3 sites, 3 technology platforms: Antibody (Freiburg, Germany) Plasmid (Fargo, USA) Protein (Madison, USA) 130 employees 1,000s of clients, including 16 of the world s top 20 pharma companies COMPANY 2

3 Aldevron Freiburg: antibodies GENERATE ANTIBODIES FROM: Directly start from an electronic cdna sequence Protein Peptide Generate highly specific antibodies (nm-pm) recognising conformational epitopes cdna Genetic immunisation: our core technology Optimised screening system against native proteins Applications: tools, diagnostics or therapeutics 18 years experience > 200 references >1700 projects 84% success rate Fee-forservice TECHNOLOGY 3

4 Benefits of GENOVAC Technology GENOVAC Genetic Immunisation Protein immunisation Peptide immunisation Antigen production Economic, faster and easier to manipulate More expensive, longer and difficult to manipulate Economic, faster Antigen characteristics Native conformation, correct folding and glycosylation Protein quality variable Linear epitopes Immune response Highly specific, no risk of contamination Risk of contaminating proteins Risk of cross reactivity Affinity High affinity (pm) Moderate to high Moderate Screening on native proteins Identification of functional mabs GENETIC IMMUNISATION 4

5 Distinguish highly conserved molecules Fofana I et al (2013), J Virol CLAUDIN Demonstrates sensitivity of the technology Differentiate small features in the extracellular loops geo mean (FACScan) 3 amino 1200 acid 800 differences in the extracellular loops between CLDN6 & CLDN9 geo mean (FACScan) Generation of mabs that differentiates two 200 comm. CLDN6 highly conserved tetraspanin molecules 0 Development of monoclonal antibodies that differentiate two highly conserved teraspanin molecules (CLDN6 & CLDN9) Development of monoclonal Development Development Development antibodies of monoclonal of monoclonal that of monoclonal differentiate antibodies antibodies that diff tha two highly conserved two highly teraspanin two highly two conserved highly conserved molecules conserved teraspanin (CLDN6 teraspanin teraspanin molecules & CLDN9) molecules (CLDN ( WU-3C9-B11 comm. CLDN6 geo mean (FACScan) geo mean (FACScan) WU-8F5-E7 WU-3C9-B geo mean (FACScan) WU-9E1-G2 0 WU-8F5-E7 0 comm. CLDN WU-10A4-B9 WU-9E1-G2 WU-10B2-A5 0 WU-10A4-B9 WU-3C9-B11 comm. CLDN6 YD-1C4-A3 mabs Commercial anti-cldn6 WU-8F5-E7 WU-10B2-A5 WU-3C9-B11 comm. CLDN6 Aldevron anti-cldn6-01 YD-1C4-A3 WU-8F5-E7 WU-9E1-G2 WU-3C9-B11 mabs YD-4E9-A2 YD-8H2-D7 Aldevron anti-cldn6-02 YD-4E9-A2 WU-8F5-E7 WU-9E1-G2 WU-10A4-B9 YD-9H8-A6 Aldevron anti-cldn6-03 YD-8H2-D7 WU-9E1-G2 WU-10A4-B9 WU-10B2-A5 WY-1H8-B7 Aldevron anti-cldn9-01 YD-9H8-A6 YD-1C4-A3 WU-10A4-B9 WU-10B2-A5 CLDN6 CLDN9 Control antigen Aldevron anti-cldn9-02 YD-4E9-A2 YD-1C4-A3 WY-1H8-B7 WU-10B2-A5 CLDN6 CLDN9 Control antigen OM-7D3-B3 Negative control YD-4E9-A2 YD-1C4-A3 YD-8H2-D7 OM-7D3-B3 mabs mabs mabs YD-4E9-A YD-9H8-A YD-8H2-D GENETIC IMMUNISATION 5

6 Agonistic mabs against a GPCR Hutchings C. (2014) mabs Mab1 and Mab3: -stimulate camp production -cross-react with turkey, rat and human β1ar -recognise different epitopes (conformational vs linear epitopes). Data not shown. -have agonistic activity Mab3 up-regulates heart rate without affecting blood pressure. Generation of monoclonal antibody that stimulates function of β1ar. Treat Bradycardia patients? β1ar mab 3 Isotype control CARDIOVASCULAR 6

7 Immunostimulatory mabs Curti B. et al. (2013), Cancer Res. Binding OX40 with agonistic anti-ox40 mabs generated by GENOVAC OX40 has been shown to inhibit apoptosis in T-cells, causing T H/Cyt -cell proliferation, antagonising T reg -cells, amplifying immune attack on tumours. MEDIMMUNE is now developing OX40 agonists for cancer therapy. Ongoing clinical phases 1 and 2. Oncolytic viruses Figure 1: Amplifying the effectivity of OX40 immunostimulation to combat tumours by applying combination therapies with e.g. checkpoint blockade inhibitors, chemotherapy, radiotherapy or armed oncolytic viruses. (modified after: Linch et al. (2015) Front. Oncol.) ONCOLOGY 7

8 Diagnostic / Therapeutic area Research tools Bacteriology Biomarker Circulation, Cardiovascular Diagnostics Immunology Immunosuppression Inflammation Neurology Oncology Degenerative bone disease Pregnancy Respiratory diseases Virology Targeted-proteins BoNT, CEACAM6, CEACAM1, CMG2, VPAC1 CD34, CEACAM5, CEACAM20, LGR4, LGR5, PDX1, SOX17, VceC, βig- H3, EphA2, CASD1 AP-FXIII, CrT, EpoR, VEGFR-2, β1ar Bouganin, Epcam, HER2, Nicastrin, S100A12 DNP, Granzyme A, K and M, HUD, NGK2D, ST2L, VPAC1 CEACAM6, FPR2, S100A4 Annexin-1, CEACAM6, FPRL1, GPR55, Granzyme K, S100A12 CEACAM16, Huntingtin, PCDHα4, EphA2 CEACAM1, CEACAM5, CEACAM6, CEACAM7, CEACAM20, CXCR7, Flamingo, IL-13R, Metaring, NGK2D, Nifie 14, OX40, FPR2, GPR30, S100A4, Shrew-1, TEM7, βig-h3, Nicastrin, DDR1b, Nectin 2, TEM1, TEM9, TEM17, RAS DKK1, Sclerostin, Wise PSG IL-13Rα1 AdE1a, CD81, CEACAM1, Claudin1, CXCR7, HAV-2B, Hendra virus F & G, HIV-GP120, HIV-GP140, Influenza A-HA, Nipah virus F & G, ScaRB1, swat, swtmprss2 8

9 Humanised mabs for Therapy Generation of humanised monoclonal antibodies using GENOVAC Antibody Technology in wild-type rats THERAPEUTICS 9

10 Humanised mabs for therapy DaTaMabs Humanisation expertise + Aldevron labs and antibody expertise = HUMANISED mabs TECHNOLOGY: Clinically validated CDR-grafting technology (FTO) - Intelligent design strategies - retain Mab potency and specificity - Identify key Framework (FW) residues and explore FW region diversity - No CDR-based mutagenesis potential solubility/aggregation issues EXPERIENCE: - 40 humanised antibodies over 25 years - 8 currently in clinical trials - 4 already on the market: Tysabri, Actemra, Entyvio & Keytruda THERAPEUTICS 10

11 Human mabs for Therapy Generation of human monoclonal antibodies using GENOVAC Antibody Technology in OMT transgenic rodents THERAPEUTICS 11

12 Protein Complex: I OBJECTIVE: Generate monoclonal antibodies in OmniRats against human TCR-complex with cross reactivity to cynomolgus TCR-complex. METHOD: Genetic Immunisation of OmniRats by co-immunisation with CD3 chains. RESULT: Successfully generated cross-reactive monoclonal antibodies tested on transfected cells (human and cyno), and on Jurkat cells. Confirmed by independent laboratories. The TCR complex is composed of 6 different polypeptides on the surface of T-cells: alpha & beta fragments CD3 gamma, delta, epsilon Zeta chains THERAPEUTICS 12

13 Protein Complex: II Three OmniRats used for two fusions yielded: positive hits out of 1056 tested samples (11%) from 2 animals positive hits out of 440 tested samples (27%) from 1 animal Initial screening performed on transfected cells. Example of one selected monoclonal antibody tested on Jurkat cells: CD3- (J.RT3-T3.5) (red) and CD3+ (E6-1) (blue). THERAPEUTICS 13

14 Established Collaboration - OMT animals housed in Germany at an animal facility approved by OMT - Breeding of OmniRats started in Multiple projects with OmniRats, OmniMouse, OmniFlic successfully applied - Various immunisation protocols successfully completed on OMT animals - Established work flow between the client, OMT, breeding facility and Aldevron from ordering and importing genetically modified animals to immunisation, fusion and screening THERAPEUTICS 14

15 Thank you Dr John Thompson +49 (0)