Phagotopes derived by antibody screening of phage-displayed random peptide libraries vary in immunoreactivity: Studies using an exemplary monoclonal antibody, CII-C1, to type II collagen

Davies, J. M., Rowley, M. J., & Mackay, I. R. (1999) Phagotopes derived by antibody screening of phage-displayed random peptide libraries vary in immunoreactivity: Studies using an exemplary monoclonal antibody, CII-C1, to type II collagen. Immunology and Cell Biology, 77(6), pp. 483-490.

View at publisher

Abstract

Antibody screening of phage-displayed random peptide libraries to identify mimotopes of conformational epitopes is promising. However, because interpretations can be difficult, an exemplary system has been used in the present study to investigate whether variation in the peptide sequences of selected phagotopes corresponded with variation in immunoreactivity. The phagotopes, derived using a well-characterized monoclonal antibody, CII-C1, to a known conformational epitope on type II collagen, C1, were tested by direct and inhibition ELISA for reactivity with CII-C1. A multiple sequence alignment algorithm, PILEUP, was used to sort the peptides expressed by the phagotopes into clusters. A model was prepared of the C1 epitope on type II collagen. The 12 selected phagotopes reacted with CII-C1 by both direct ELISA (titres from < 100-11 200) and inhibition ELISA (20-100% inhibition); the reactivity varied according to the peptide sequence and assay format. The differences in reactivity between the phagotopes were mostly in accord with the alignment, by PILEUP, of the peptide sequences. The finding that the phagotopes functionally mimicked the C1 epitope on collagen was validated in that amino acids RRL at the amino terminal of many of the peptides were topographically demonstrable on the model of the C1 epitope. Notably, one phagotope that expressed the widely divergent peptide C-IAPKRHNSA-C also mimicked the C1 epitope, as judged by reactivity in each of the assays used: these included cross-inhibition of CII-C1 reactivity with each of the other phagotopes and inhibition by a synthetic peptide corresponding to that expressed by the most frequently selected phagotope, RRLPFGSQM. Thus, it has been demonstrated that multiple phage-displayed peptides can mimic the same epitope and that observed immunoreactivity of selected phagotopes with the selecting mAb can depend on the primary sequence of the expressed peptide and also on the assay format used.

Impact and interest:

10 citations in Scopus
Search Google Scholar™
10 citations in Web of Science®

Citation counts are sourced monthly from Scopus and Web of Science® citation databases.

These databases contain citations from different subsets of available publications and different time periods and thus the citation count from each is usually different. Some works are not in either database and no count is displayed. Scopus includes citations from articles published in 1996 onwards, and Web of Science® generally from 1980 onwards.

Citations counts from the Google Scholar™ indexing service can be viewed at the linked Google Scholar™ search.

ID Code: 87930
Item Type: Journal Article
Refereed: Yes
Keywords: Monoclonal antibody, Phage display, Random peptide libraries, Type II collagen, collagen antibody, collagen type 2, epitope, peptide library, amino acid sequence, amino terminal sequence, article, controlled study, enzyme linked immunosorbent assay, immunoreactivity, Animals, Antibodies, Monoclonal, Antibody Specificity, Antigen-Antibody Reactions, Binding, Competitive, Cattle, Collagen, Combinatorial Chemistry Techniques, Enzyme-Linked Immunosorbent Assay, Epitopes, Models, Molecular, Peptides, Serologic Tests
DOI: 10.1046/j.1440-1711.1999.00846.x
ISSN: 08189641
Divisions: Current > Schools > School of Biomedical Sciences
Current > QUT Faculties and Divisions > Faculty of Health
Copyright Owner: © 1999 Australasian Society for Immunology
Deposited On: 01 Oct 2015 02:40
Last Modified: 01 Oct 2015 02:40

Export: EndNote | Dublin Core | BibTeX

Repository Staff Only: item control page