Enhancing thermal stability in the CH₂ domain to suppress aggregation through the introduction of simultaneous disulfide bonds in Pichia pastoris.

ABSTRACT: Protein aggregations decrease production yields and impair the efficacy of therapeutics. The CH₂ domain is a crucial part of the constant region of human IgG. But, it is also the least stable domain in IgG, which can result in antibody instability and aggregation problems. We created a novel mutant of the CH₂ domain (T250C/L314C, mut10) by introducing a disulfide bond and expressed it using Pichia pastoris. The mut10 variant exhibited enhanced thermal stability, resistance to enzymatic degradation, and reduced aggregation in comparison to the original CH₂ domain. However, it was less stable than mut20 (L242C/K334C), which is the variant prepared in a previous study (Gong et al., J. Biol. Chem., 2009). A more advanced mutant, mut25, was created by combining

SUBMITTER: Oyama K 

PROVIDER: S-EPMC10680342 | biostudies-literature | 2023 Dec

REPOSITORIES: biostudies-literature