Characterization of the internal translation initiation region in monoclonal antibodies expressed in Escherichia coli.

ABSTRACT: Monoclonal antibodies (mAbs) represent an important platform for the development of biotherapeutic products. Most mAbs are produced in mammalian cells, but several mAbs are made in Escherichia coli, including therapeutic fragments. The NISTmAb is a well-characterized reference material made widely available to facilitate the development of both originator biologics and biosimilars. Here, when expressing NISTmAb from codon-optimized constructs in E. coli (eNISTmAb), a truncated variant of its heavy chain was observed. N-terminal protein sequencing and mutagenesis analyses indicated that the truncation resulted from an internal translation initiation from a GTG codon (encoding Val) within eNISTmAb. Using computational and biochemical approaches, we demonstrate that this transla

SUBMITTER: Leith EM 

PROVIDER: S-EPMC6885640 | biostudies-literature | 2019 Nov

REPOSITORIES: biostudies-literature