C-terminal antibodies (CTAbs): a simple and broadly applicable approach for the rapid generation of protein-specific antibodies with predefined specificity.
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.
Edwards
Robert J
RJ
Experimental Medicine and Toxicology, Imperial College London, London, UK. r.edwards@imperial.ac.uk
Wrigley
Amanda
A
Bai
Zhonghu
Z
Bateman
Michaela
M
Russell
Hugh
H
Murray
Stephen
S
Lu
Huafeng
H
Taylor
Graham W
GW
Boobis
Alan R
AR
Sriskandan
Shiranee
S
D000595:Amino Acid Sequence; D000906:Antibodies / Q000276:immunology*; D000918:Antibody Specificity; D001426:Bacterial Proteins / Q000276:immunology*; D013297:Streptococcus pyogenes / Q000276:immunology / Q000472:pathogenicity
D000906:Antibodies; D001426:Bacterial Proteins