Monoclonal antibody MAb M1169 identifies surface proteins of thermotolerant campylobacter species
LE3 .A278 2017
Bachelor of Science
Campylobacter spp. is a Gram negative, thermotolerant bacterium and a significant foodborne pathogen. Transmission is mainly through ingestion of undercooked and ready-to-eat poultry and mishandling of contaminated foods. While most individuals who have acquired campylobacteriosis recover rapidly (seven to ten days), individuals who are immunocompromised may develop rare, and potentially lethal complications, such as Guillain-Barre Syndrome. Recently, a colony blot immunoassay has been developed at the Canadian Food Inspection Agency (CFIA) using a Monoclonal Antibody (MAb) M1169 M1169 for the detection of thermotolerant Campylobacter spp. In this study, we further characterized the surface target proteins of C. jejuniand confirmed specificity of this MAb to thermotolerant Campylobacter spp. using Western blot analysis. After immunoprecipitation, M1169 successfully detected only thermotolerant Campylobacter spp. by Western blotting analysis. Multiple protein bands were reacted with M1169, and protein bands were excised from 1D SDS- PAGE and analyzed using mass spectrometry (MS). The main proteins that were identified included major outer membrane protein (OMP) and flagellin. Other proteins included cytochrome C oxidase which is a large integral membrane protein found in prokaryotes. This work will help in advancing methods used in the detection and identification of C. jejuniin Canadian poultry, and in the development of a rapid detection method for thermotolerant Campylobacter spp. for CFIA.
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