Identification of a specific surface epitope of OmpC for Escherichia coli O157:H7 with protein topology facilitated affinity mass spectrometry.

ABSTRACT: The goal of this work was to identify the target protein and epitope of a previously reported Escherichia coli O157:H7 (ECO157)-specific monoclonal antibody (mAb) 2G12. mAb 2G12 has shown high specificity for the recovery and detection of ECO157. To achieve this goal, the target protein was first separated by two-dimensional gel electrophoresis (2-DE) and located by Western blot (WB). The protein spots were identified to be the outer membrane protein (Omp) C by matrix-assisted laser desorption/ionization time-of-flight mass spectrometry (MALDI-TOF-MS). After that, the target protein was purified by immunoaffinity chromatography (IAC) and subjected to in situ enzymatic cleavage of the vulnerable peptides. Eight eluted peptides of OmpC identified by liquid chromatography-tandem mass spectrom

SUBMITTER: Wang W 

PROVIDER: S-EPMC8426304 | biostudies-literature | 2021 Sep

REPOSITORIES: biostudies-literature