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