Copper assisted sequence-specific chemical protein conjugation at a single backbone amide.

ABSTRACT: Direct, site-specific methods of protein functionalization are highly desirable for biotechnology. However, such methods are challenging due to the difficulty of chemically differentiating a single site within a large protein. Herein, we propose "metal binding targeting" strategy and develop a Copper Assisted Sequence-specific conjugation Tag (CAST) method to achieve rapid (second order rate 8.1 M^-1 s^-1), site-specific protein backbone chemical modification with pinpoint accuracy. We demonstrate the versatility of CAST conjugation by preparing various on-demand modified recombinant proteins, including a homogeneous antibody-drug conjugate with high plasma stability and potent efficacy in vitro and in vivo. Thus, CAST provides an efficient and quantitative method to si

SUBMITTER: Guo M 

PROVIDER: S-EPMC10698186 | biostudies-literature | 2023 Dec

REPOSITORIES: biostudies-literature