Interdomain Interactions Modulate Refolding Kinetics and Aggregation in a Monoclonal Antibody.

ABSTRACT: Understanding the structural determinants of antibody stability and aggregation is essential for therapeutic development. In this study, we investigated the unfolding and refolding behavior of bevacizumab under denaturing conditions using dynamic light scattering (DLS), circular dichroism (CD), and hydrogen-deuterium exchange mass spectrometry (HDX-MS). Unfolding was induced by incubating the antibody in 4 M guanidine hydrochloride (Gnd-HCl), followed by refolding through dilution with 1 M Gnd-HCl. Each domain exhibited distinct unfolding kinetics: the C_H2 and V_H domains unfolded rapidly, while the C_H3 domain retained its structure until 45 min, consistent with its known thermodynamic stability. Aggregation, detected by DLS, was prevalent only after 120 min

SUBMITTER: Trolese P 

PROVIDER: S-EPMC12598857 | biostudies-literature | 2025 Nov

REPOSITORIES: biostudies-literature