Open AccessCharacterisation of a monoclonal antibody and its use in the immunoaffinity purification of penicillin‐binding protein 2′ of methicillin‐resistant Staphylococcus aureus
Author(s) -
Harrington Charles R.,
O'Hara Denise M.,
Reynolds Peter E.
Publication year - 1989
Publication title -
fems microbiology letters
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.899
H-Index - 151
eISSN - 1574-6968
pISSN - 0378-1097
DOI - 10.1111/j.1574-6968.1989.tb03612.x
Subject(s) - monoclonal antibody , staphylococcus aureus , affinity chromatography , antibody , protein a , sepharose , penicillin binding proteins , microbiology and biotechnology , biology , immunoprecipitation , antigen , chemistry , penicillin , biochemistry , bacteria , antibiotics , enzyme , immunology , genetics
The additional penicillin‐binding protein (PBP) 2′ that is important in determining intrinsic resistance in methicillin‐resistant strains of Staphylococcus aureus (MRSA) has been purified by affinity chromatography using monoclonal antibodies. Monoclonal antibody 1/423.10.351 reacted in ELISA with detergent extracts of membranes from resistant organisms, but not in immunoblots with PBP 2′ separated by SDS‐PAGE. Immunoprecipitation experiments showed that antibody 1/423.10.351 reacted with PBP 2′ in detergent extracts. The latter antibody, covalently coupled to protein A‐Sepharose through the Fc region, served as an affinity matrix to purify PBP 2′. The PBP was detected in immunoblots using a second monoclonal antibody, 2/401.43. Conjugation of this antibody with alkaline phosphatase afforded more rapid detection of PBP 2′ for the immunological detection of PBP 2′ both in affinity‐purified fractions and in resistant strains.