Immunodiscovery developing antibodies for research, diagnosis and therapy MATS OHLIN

Transcription

1 Immunodiscovery developing antibodies for research, diagnosis and therapy MATS OHLIN

2 Immunotechnology a department at Lund University exploiting advanced technologies in biomedicine Genomics Antibody technology Immunotechnology Proteomics Antibody-driven proteomics Allergy Cancer Immunology Bioinformatics

3 Antibodies key molecules exploiting advanced technologies in biomedicine Genomics Antibody technology Immunotechnology Proteomics Antibody-driven proteomics Allergy Cancer Immunology Bioinformatics

4 The versatile antibody Target binding paratope 90 Effector function Fc

5 Targets of antibodies Antigens and their epitopes Kwong and Mascola (2012) Cell 37,

6 The target antigen A diagnostic molecule

7 Wingren et al. (2012) Cancer Res 72, 2481 Antibody-based discovery of biomarkers Multiplexed diagnostic or prognostic sets of analytes

8 The target antigen Diagnostic molecules A research target A cellular component for therapy Presence on target cell Unwanted presence on other cells (off-target effects) Accessibility Antibody mechanism of action

9 Diverse functions in oncology Weiner (2015) Nat Rev Cancer 15,

10 Biological function of antibodies mechanisms of action Agonist / antagonist / partial antagonist Steric hindrance of activity Prevention of dimerization Signaling Induction of apoptosis Complement activation Opsonization Antibody-dependent cellular cytotoxicity Carrier payload Anti-angiogenesis Activation of immunity

11 Antibodies as registered drugs

12 Recombinant antibodies $75 billion annual sales (2013) * * * anti-vegf IFN-β1a rtnfr anti-her2 anti-tnfα IFN-β1a anti-tnfα anti-cd20 Ecker et al. (2015) mabs 7, 9-14

13 Targets and indications

14 Immune checkpoint regulators new fancy targets

15 Immune checkpoint regulators new fancy targets controlling immune activity

16 The binder (Antisera) Mouse monoclonal antibody

17 The foreign nature of mouse antibodies The problem immunogenicity of non-human proteins the difficulty of raising human antibodies Mouse antibody sequence

18 The binder solving the problem through technology advances (Antisera) Mouse monoclonal antibody Chimeric antibody Humanized antibody Human antibody From human immune response From mice with human antibody locus From combinatorial antibody library

19 Chimeric and humanized antibodies enhancing the proteins human nature The problems immunogenicity of non-human proteins and the difficulty of raising human antibodies Mouse antibody Chimeric antibody Humanized antibody Human antibody sequence sequence sequence sequence Nature 1986; 321, 522

20 A diverse set of constructs

21 Harvesting human monoclonal antibody specificities from immune individuals B cell immortalization Human hybridoma technology Antibody gene cloning Blood collection Single cell sorting Gene cloning Antibody product analysis

22 Jakobovits et al. (2007) Nat Biotechnol 25, Mice producing human antibodies

23 Antibodies from combinatorial libraries Collections of millions billions of antibody encoding genes and their corresponding antibody specificities From immune response Naïve development of antibodies against any target

24 Phage display technology Screen for desired properties Immobilized Antigen 4 3 Immobilized Antigen 1. Immobilize antigen 2. Add phage library 3. Wash away non-binding phages 4. Elute bound phages and infect E.coli 5. Amplify binding phages 6. Repeat step 1-5 until sufficient enrichment 7. Screen for desired properties

25 Phage display technology the evolution approach * * * * * * * * * * * * * * Immobilized Antigen Immobilized Antigen

26 Amstutz et al. (2001) Curr Opin Biotechnol 12, Ribosomal display Very large libraries Built-in protein evolution

27 Large-scale antibody development pipeline Large library Automated selection Automated production High throughput immunoassay / bioassays Designed plasmids for ease of cloning Prokaryotic and eukaryotic cell production systems

28 Antibody formats Intact naked antibodies choice of isotype (e.g. IgG1-4), fragment etc. Single antibody vs. antibody mixtures Immunoconjugates CAR T cells

29 Example: CD20 as a target Antibody: Rituximab Product: MabThera Target: present on most cells of the B cell lineage Treatment of disease with B cell involvement e.g. Non-Hodgkin s lymphoma & chronic lymphatic leukemia Rheumatoid arthritis Granulomatosis with polyangiitis and microscopic polyangiitis Antibody depletes B cells Impairs antibody responses Antibody does not affect plasma cells or stem cells Does not eliminate existing antibody production

30 Immunoconjugates Kadcyla targeting HER2+ cells with chemotherapy (Emtansine)

31 CAR-T cells and bispecific antibodies adding function to targeting Recruiting T cell immunity to tumors Suzuki et al. (2015) Ped Blood Cancer 62,

32 Biosimilar a biological drug that is almost an identical copy of an original product

33 Discovery in the field of antibody A critical source of new entities for biomedicine and research

34