Pre-Clinical In-Vitro Studies on Parameters Governing Immune Complex Formation

Area of Science:

• Biopharmaceutical development
• Immunology
• Analytical chemistry

Background:

• Biotherapeutic success is hindered by immunogenicity, leading to anti-drug antibody (ADA) formation.
• ADAs can form immune complexes (ICs) with biotherapeutics, causing downstream cellular effects.
• The relationship between IC characteristics and their biological impact remains unclear.

Purpose of the Study:

• To systematically investigate factors influencing in vitro immune complex formation.
• To characterize immune complex species and their properties.
• To provide insights into biotherapeutic immunogenicity and risk assessment.

Main Methods:

• Utilized classic analytical techniques: Size Exclusion Chromatography-Multi-Angle Light Scattering (SEC-MALS) and Sedimentation Velocity Analytical Ultracentrifugation (SV-AUC).
• Employed advanced mass photometry for state-of-the-art characterization.
• Designed a surrogate study to assess IC formation dynamics.

Main Results:

• Demonstrated a significant interplay between component concentrations and molar ratios in IC formation.
• Highlighted the influence of the biologic's and its target's structural features on IC characteristics.
• Validated a versatile analytical toolbox for IC assessment.

Conclusions:

• Established that component concentrations, molar ratios, and structural features critically govern IC formation and properties.
• The developed methodology is applicable to various biotherapeutics for early immunogenicity risk detection.
• Enables evaluation of how degraded or modified biotherapeutics impact IC formation.