Balancing the extremes for antibody developability: hydrophobic and electrostatic germline framework signatures for...

Area of Science:

• Biopharmaceutical development
• Protein engineering
• Immunology

Background:

• Antibody therapeutics are crucial but face developability challenges.
• Complementarity-determining region (CDR) modifications impact antigen specificity.
• Framework regions offer opportunities to improve developability attributes like aggregation and viscosity.

Purpose of the Study:

• To systematically analyze the electrostatic and hydrophobic surface properties of germline antibody frameworks.
• To assess the role of these properties in modulating antibody fragment variable (Fv) developability.
• To demonstrate how framework selection can optimize antibody developability.

Main Methods:

• Structure prediction and surface patch analysis of germline antibody frameworks.
• Characterization of kappa and lambda light-chain frameworks.
• Hydrophobic interaction chromatography and heparin column analysis.
• Assessment of framework selection's impact on physicochemical characteristics.

Main Results:

• Identified differences in surface properties between kappa and lambda light-chain frameworks.
• Characterized outlier germlines with extreme surface properties.
• Demonstrated that framework selection can compensate for CDR loop physicochemical properties.
• Showcased framework selection as a tool to optimize antibody developability.

Conclusions:

• Rational framework selection is a systematic approach to enhance antibody developability.
• Leveraging inherent germline properties provides a toolbox for antibody design.
• This strategy improves therapeutic candidate selection for biopharmaceuticals.