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Prediction of Aggregation Propensity and Monitoring of Aggregation of Antibody-Drug Conjugates (ADC) using ProteoStat® Reagents

Nicolas Schneider1, Morgan Mathieu2, Sandra Savard-Chambard Gas1, Angélique Boedec Herbette1, Hélène Rispaud1, Naouel Lovera1, Delphine Bregeon1, Agnès Represa1, Christian Belmant1
1 Innate Pharma, Marseille, France
2 Enzo Life Sciences, Lausen, Switzerland

Featured Product: ProteoStat® Protein Aggregation Standards (IgG), ProteoStat® Protein Aggregation Assay, ProteoStat® Thermal Shift Stability Assay

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Abstract

Innate Pharma has developed an antibody-drug conjugate (ADC) coupling technology. The stability of unglycosylated SGN30 Q and S mutants coupled to the MMAE toxin by either one-step (SGN30 Q/S-linker-vcMMAE) or two-step technology (SGN30 Q/S-Linker-Click-Linker-vcMMAE) was compared to the first FDA-approved ADC, ADCETRIS®, and evaluated to validate the technology concept for clinical applications. The temperatures of aggregation, determined via the ProteoStat® Thermal Shift Assay, established the following aggregation propensity prediction ladder: SGN30 Q ADCs coupled with four MMAE toxins are less stable than SGN30 S ADCs coupled with only two MMAE toxins; ADCs coupled with lipophilic linkers are less stable than ADCs coupled with hydrophilic ones; and ADCETRIS® is more stable than a SGN30 Q ADC coupled to a lipophilic linker but less stable than a SGN30 Q ADC coupled with a hydrophilic one. An accelerated degradation study confirmed this hierarchy (with the exception of ADCETRIS® likely due to a higher fragmentation of its antibody element) and provided confirmation that the ProteoStat® Thermal Shift Assay is predictive of aggregation propensity for ADC products.

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