Fast protein sequencing of monoclonal antibody by real-time digestion on emitter during nanoelectrospray.

ABSTRACT: Growth in the pharmaceutical industry has led to an increasing demand for rapid characterization of therapeutic monoclonal antibodies. The current methods for antibody sequence confirmation (e.g., N-terminal Edman sequencing and traditional peptide mapping methods) are not sufficient; thus, we developed a fast method for sequencing recombinant monoclonal antibodies using a novel digestion-on-emitter technology. Using this method, a monoclonal antibody can be denatured, reduced, digested, and sequenced in less than an hour. High throughput and satisfactory protein sequence coverage were achieved by using a non-specific protease from Aspergillus saitoi, protease XIII, to digest the denatured and reduced monoclonal antibody on an electrospray emitter, while electrospray high voltage was appli

SUBMITTER: Mao Y 

PROVIDER: S-EPMC6601538 | biostudies-literature | 2019 May/Jun

REPOSITORIES: biostudies-literature