Hinge Truncation to Improve Aggregation Kinetics and Thermal Stability of an Antibody Fab Fragment.

ABSTRACT: The hinge region of antibody fragments plays a crucial role in their stability and aggregation properties. In this study, we investigated the effects of hinge truncations on the thermal stability and aggregation propensity of the A33 Fab antibody fragment. Eight Fab variants were engineered by introducing stop codons to truncate 1-8 residues at the hinge region (heavy chain residues 221-228). These variants were then expressed, purified, and characterized in terms of stability and aggregation propensity using SDS-PAGE, SEC-HPLC, LC-MS, and thermal stability assays. Our findings demonstrate that truncating the hinge region can enhance the thermal stability and reduce the aggregation of Fab fragments, and that progressive truncations identified an optimal hinge length for stability. Notably,

SUBMITTER: Zhang C 

PROVIDER: S-EPMC12406249 | biostudies-literature | 2025 Sep

REPOSITORIES: biostudies-literature