Expression and Characterization of Intein-Cyclized Trimer of Staphylococcus aureus Protein A Domain Z.

ABSTRACT: Staphylococcus aureus protein A (SpA) is an IgG Fc-binding virulence factor that is widely used in antibody purification and as a scaffold to develop affinity molecules. A cyclized SpA Z domain could offer exopeptidase resistance, reduced chromatographic ligand leaching after single-site endopeptidase cleavage, and enhanced IgG binding properties by preorganization, potentially reducing conformational entropy loss upon binding. In this work, a Z domain trimer (Z3) was cyclized using protein intein splicing. Interactions of cyclic and linear Z3 with human IgG₁ were characterized by differential scanning fluorimetry (DSF), surface plasmon resonance (SPR), and isothermal titration calorimetry (ITC). DSF showed a 5 ℃ increase in IgG₁ melting temperature when bound

SUBMITTER: Nandy S 

PROVIDER: S-EPMC9865183 | biostudies-literature | 2023 Jan

REPOSITORIES: biostudies-literature