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C‐Terminal antibodies (CTAbs): A simple and broadly applicable approach for the rapid generation of protein‐specific antibodies with predefined specificity
Author(s) -
Edwards Robert J.,
Wrigley Amanda,
Bai Zhonghu,
Bateman Michaela,
Russell Hugh,
Murray Stephen,
Lu Huafeng,
Taylor Graham W.,
Boobis Alan R.,
Sriskandan Shiranee
Publication year - 2007
Publication title -
proteomics
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.26
H-Index - 167
eISSN - 1615-9861
pISSN - 1615-9853
DOI - 10.1002/pmic.200600916
Subject(s) - immunogen , antibody , computational biology , streptococcus pyogenes , biology , protein engineering , proteomics , biochemistry , peptide sequence , chemistry , bacteria , monoclonal antibody , immunology , genetics , gene , enzyme , staphylococcus aureus
Recent advances in proteomic techniques have resulted in an ever‐increasing need to produce antibodies. Here, to address this problem, a technically simple approach of targeting the extreme C‐termini of proteins with antibodies (CTAbs) was investigated in proteins secreted by the human pathogen Streptococcus pyogenes . Target proteins were identified by a conventional proteomic approach and CTAbs produced against synthetic five amino acid peptides representing the C‐terminus of each target protein. In every case where protein secretion was demonstrated ( n = 20), CTAbs were successfully produced and bound specifically to the target protein (100% success rate). The apparent specificity was consistent with the structural heterogeneity of the C‐termini of S. pyogenes proteins. The global specificity of CTAb binding was defined using a combinatorial library of synthetic peptides representing structural variants of the original synthetic immunogen. This is a systematic and comprehensive approach for the development of antibodies with defined specificity that can be used in a range of applications.