Slide 1: Engineering a Pure and Stable Heterodimeric IgA for Multispecific Therapeutics
Engineering a Pure and Stable Heterodimeric IgA for the Development of Multispecific Therapeutics PepTalk 2024 Meghan Verstraete, PhD Nasdaq: ZYME | zymeworks.com
Slide 2: Legal Disclaimer
Legal Disclaimer This presentation includes “forward-looking statements” within the meaning of the U.S. Private Securities Litigation Reform Act of 1995 and “forward-looking information” within the meaning of Canadian securities laws, or collectively, forward-looking statements. Forward-looking stat
Slide 3: Advances in IgG Protein Engineering Fueled Multispecific Antibody Development
Advances in IgG Protein Engineering has Fueled Rapid Development and Expansion of Multispecific Antibody Formats and Their Applications Bispecific IgG antibodies have been developed More than 30 mature commercial technology with broad application for cancer immunotherapy platforms have been used to
Slide 4: IgG is not the Only Antibody Isotype that can be Leveraged
IgG is not the Only Antibody Isotype that can be Leveraged for Therapeutic Design IgM for avidity and Fc-mediated IgA for Fc-mediated effector functions IgE for Fc-mediated effector functions effector functions (eg. complement) (eg. neutrophil activation) (eg. macrophage repolarization) Engineering
Slide 5: Recent Work has Highlighted the Potential of IgA as a Cancer Therapeutic
Recent Work has Highlighted the Potential of IgA as a Cancer Therapeutic • Neutrophils are a large untapped • Utility of IgA to engage neutrophils • IgA can potently activate effector cell population that could in vitro and in vivo to kill cancer cells neutrophils via binding to the be harnessed to
Slide 6: Engineering for Developability: Previous Success with IgG Heterodimeric Fc
Engineering for Developability: Zymeworks’ Previous Success Engineering an IgG Heterodimeric Fc Developability concepts and parameters for purity and stability introduced early in the engineering process Azymetric is a best-in-class IgG heterodimeric antibody technology developed using proprietary i
Slide 7: Rational Design Strategy Used for IgA Fc Interface Engineering
Rational Design Strategy Used for IgA Fc Interface Engineering Engineer for Engineer for heterodimer heterodimer purity using stability using negative design positive design 1. Structure Model 5. Experimental 2. System Analysis 3. In silico design 4. Variant selection Creation screening Making a Mea
Slide 8: Crystal Structure of Homodimeric IgA Fc Used for In-depth Analysis
Crystal Structure of Homodimeric (Wild-type) IgA Fc Used for in- depth Analysis of IgA CH3-CH3 Interface 1. Structure Model IgA Fc interface contact analysis 2. System Analysis Creation IgA Fc chain A IgA Fc chain B IgA Fc CH2 Fc⍺RI Fc⍺RI PDB ID:1OW0; from Herr et al, Nature, 2003 1. Interface analy
Slide 9: In Silico Tools Employed to Inform Design
In Silico Tools Employed to Inform Heterodimeric IgA Fc Design 3. In silico design Variants were predominately selected based on affinity metrics Negative designs Models of homodimeric and heterodimeric IgA CH3:CH3 (“breaking” homodimerization) variants were scored for affinity based on: Predicted a
Slide 10: Example In Silico Models of Steric vs Electrostatic Designs
Example In Silico Models of Steric vs Electrostatic Heterodimeric IgA Fc Designs 4. Variant selection W398T A412F I416E L372R L396 V T414 Y T366D I416L Top 12 designs selected for experimental screening Making a Meaningful Difference 10
Slide 11: Experimental In Vitro Evaluation of IgA Heterodimeric Fc Designs
Experimental In Vitro Evaluation of IgA Heterodimeric Fc Designs One-armed antibody (OAA) format used to test top-ranked designs • Unique sets of mutations introduced into IgA CH3 domains for “chain A” and “chain B” Making a Meaningful Difference 11
Slide 12: High Purity Measured by CE-SDS and Analytical SEC
High Heterodimeric IgA Fc Purity Measured by CE-SDS and Analytical SEC after Affinity Purification 5. Experimental screening Making a Meaningful Difference 12
Slide 13: Engineered for Thermal Stability and High Purity
Heterodimeric IgA Fc Engineered for Thermal Stability and High Purity Thermal stability of purified heterodimeric IgA Fc designs assessed by differential scanning calorimetry Summary of purity and stability for IgA Fc design Post affinity purification Post prepSEC purification Design HPLC-SEC CE-SDS
Slide 14: Crystal Structure Shows Mutations Do Not Perturb IgA Fc Structure
A Crystal Structure of the Heterodimeric IgA Fc Revealed that the IgA CH3 Mutations do not Perturb the Overall IgA Fc Structure and a Heterodimeric IgA Interface Consistent with In Silico Models Heterodimeric IgA Fc Chain A mutations Chain B mutations (steric 6 design) L396V, W398T, I416L A412F, T41
Slide 15: Heterodimeric IgA Fc Designs Show Preserved Binding to FcαRI by SPR
Heterodimeric IgA Fc Designs Show Preserved Binding to Fc⍺RI by SPR Analyte: IgA Fc variant Fc⍺RI binding site Increasing the density of FcαRI resulted in Ligand: Fc⍺RI X Fc⍺RI binding site knockout (KO) higher avidity-driven binding for wild-type and heterodimeric IgA Fc variants IgA variant Varian
Slide 16: Applications of Heterodimeric IgA Fc
Applications of Heterodimeric IgA Fc Investigate IgA:Fc⍺RI IgA half-life stoichiometry required modulation for neutrophil activation Is bivalent binding of Fc⍺RI required for neutrophil activation? FcRn and Fc⍺RI binding sites are IgA structurally superimpose PDB ID: 1OW0 PDB ID: 1I1A Making a Meani
Slide 17: Applications of Heterodimeric IgA Fc: Multispecific IgA-based Formats
Applications of Heterodimeric IgA Fc Multispecific IgA- based formats Dimeric IgA specifically disables intracellular mutated oncodrivers Multispecific therapeutics targeting mucosa Figure from Prince and Hollmen, Immunity Previews, 2023 Biswas et al, Immunity, 2023 Biswas et al, Nature, 2021 Making
Slide 18: Summary
Summary Protein Engineering Multispecific IgA-based Integrating purity and formats stability design requirements yielded a heterodimeric IgA Fc with favorable developability characteristics Multispecific therapeutics Application targeting mucosa A heterodimeric IgA Fc unlocks the potential to explor
Slide 19: Acknowledgments
Acknowledgments Zymeworks team members (past and present) Florian Heinkel Eric Escobar Thomas Spreter Von Kreudenstein Surjit Dixit Siran Cao Janessa Li Patrick Farber Begonia Silva-Moreno Elizabeth Stangle University of Victoria Martin Boulanger Photo from CBRE Fangni Peng Making a Meaningful Diffe
Slide 20: Thank you
Thank you Making a Meaningful Difference 20