Comparison of Antigenic Regions Identified on IgG1Fc Using Bioinformatics vs Pepscan Analysis

2.50
Hdl Handle:
http://hdl.handle.net/2436/30232
Title:
Comparison of Antigenic Regions Identified on IgG1Fc Using Bioinformatics vs Pepscan Analysis
Authors:
Nelson, Paul N.; Westwood, Olwyn M. R.; Freimanis, Graham L.; Roden, Denise A.; Sissaoui, Samir; Rylance, Paul; Hay, Frank C.
Abstract:
Epitope mapping allowed the location of antigenic determinants on a protein macromolecule to be identified. In particular, pepscan techniques that utilize a series of overlapping peptides, help detect key amino acid residues that are important in antibody recognition and binding. In a previous study, we employed 15-mer peptides spanning the entire length of IgG1Fc to ascertain successfully the target epitopes of isotypic/allotypic monoclonal reagents. As an extension to this work we have used these peptides to evaluate the location of epitope targets of five IgM rheumatoid factor antibodies (RFAbs). Overall, 2 antibodies, RFAb TS2 and TS1, detected a similar epitope within the CH3 domain (360-KNQVSLTCLVKGFYP-374), whilst 1 (RFAb SJ1) recognised an epitope in the CH2 domain (294- EQYNSTYRVVSVLTV-308). In contrast, 2 RFAbs, PRSJ2 and PRTS1 detected four and five epitopes respectively within the Fc region. RFAb PRSJ2 recognised epitopes detected by RFAB TS2 and TS1 but also further epitopes in the CH2 domain (256-TPEVTCVVVDVSHED-270) and CH3 domain (418-QQGNVFSCSVMHEAL-432). Similarly, RFAb PRTS1 detected all four epitopes plus a fifth in the CH3 domain (382-ESNGQPENNYKTTPP-396). In essence there was a consensus of target epitopes identified by these rheumatoid factor antibodies. Interestingly, two epitopes (256–270, CH2 domain and 360–374, CH3 domain) were novel in that they had not been identified in previous pepscan studies. The other epitopes recognised, either overlapped or were immediately adjacent to previous epitopes detected by poly/monoclonal rheumatoid factor antibodies. Molecular modelling (PCImdad) of IgG1Fc showed that all five epitopes were exposed and surface accessible for antibody interaction. In addition, a bioinformatics analysis of the Fc region using ExPASy was employed to identify key antigenic determinants. This ‘in silico’ approach may provide a means of determining key regions without the need to develop overlapping peptides spanning the entire length of a macromolecule.
Citation:
Clinical Medicine: Arthritis and Musculoskeletal Disorders, 2008(1): 33-42
Publisher:
Libertas Academica Press
Journal:
Clinical Medicine: Arthritis and Musculoskeletal Disorders
Issue Date:
2008
URI:
http://hdl.handle.net/2436/30232
Additional Links:
http://la-press.com/article.php?article_id=780
Type:
Article
Language:
en
Description:
Clinical Medicine: Arthritis and Musculoskeletal Disorders is an international, open access, peer reviewed journal.
ISSN:
1178-1149
Appears in Collections:
Molecular Immunology Research Group

Full metadata record

DC FieldValue Language
dc.contributor.authorNelson, Paul N.-
dc.contributor.authorWestwood, Olwyn M. R.-
dc.contributor.authorFreimanis, Graham L.-
dc.contributor.authorRoden, Denise A.-
dc.contributor.authorSissaoui, Samir-
dc.contributor.authorRylance, Paul-
dc.contributor.authorHay, Frank C.-
dc.date.accessioned2008-06-20T09:58:59Z-
dc.date.available2008-06-20T09:58:59Z-
dc.date.issued2008-
dc.identifier.citationClinical Medicine: Arthritis and Musculoskeletal Disorders, 2008(1): 33-42en
dc.identifier.issn1178-1149-
dc.identifier.urihttp://hdl.handle.net/2436/30232-
dc.descriptionClinical Medicine: Arthritis and Musculoskeletal Disorders is an international, open access, peer reviewed journal.en
dc.description.abstractEpitope mapping allowed the location of antigenic determinants on a protein macromolecule to be identified. In particular, pepscan techniques that utilize a series of overlapping peptides, help detect key amino acid residues that are important in antibody recognition and binding. In a previous study, we employed 15-mer peptides spanning the entire length of IgG1Fc to ascertain successfully the target epitopes of isotypic/allotypic monoclonal reagents. As an extension to this work we have used these peptides to evaluate the location of epitope targets of five IgM rheumatoid factor antibodies (RFAbs). Overall, 2 antibodies, RFAb TS2 and TS1, detected a similar epitope within the CH3 domain (360-KNQVSLTCLVKGFYP-374), whilst 1 (RFAb SJ1) recognised an epitope in the CH2 domain (294- EQYNSTYRVVSVLTV-308). In contrast, 2 RFAbs, PRSJ2 and PRTS1 detected four and five epitopes respectively within the Fc region. RFAb PRSJ2 recognised epitopes detected by RFAB TS2 and TS1 but also further epitopes in the CH2 domain (256-TPEVTCVVVDVSHED-270) and CH3 domain (418-QQGNVFSCSVMHEAL-432). Similarly, RFAb PRTS1 detected all four epitopes plus a fifth in the CH3 domain (382-ESNGQPENNYKTTPP-396). In essence there was a consensus of target epitopes identified by these rheumatoid factor antibodies. Interestingly, two epitopes (256–270, CH2 domain and 360–374, CH3 domain) were novel in that they had not been identified in previous pepscan studies. The other epitopes recognised, either overlapped or were immediately adjacent to previous epitopes detected by poly/monoclonal rheumatoid factor antibodies. Molecular modelling (PCImdad) of IgG1Fc showed that all five epitopes were exposed and surface accessible for antibody interaction. In addition, a bioinformatics analysis of the Fc region using ExPASy was employed to identify key antigenic determinants. This ‘in silico’ approach may provide a means of determining key regions without the need to develop overlapping peptides spanning the entire length of a macromolecule.en
dc.language.isoenen
dc.publisherLibertas Academica Pressen
dc.relation.urlhttp://la-press.com/article.php?article_id=780en
dc.subjectEpitome mappingen
dc.subjectRheumatoid Factoren
dc.subjectBioinformaticsen
dc.subjectPeptidesen
dc.subjectMonoclonal antibodiesen
dc.subjectMolecular Biologyen
dc.subjectAntibodiesen
dc.subjectAntigensen
dc.subjectPepscan analysisen
dc.titleComparison of Antigenic Regions Identified on IgG1Fc Using Bioinformatics vs Pepscan Analysisen
dc.typeArticleen
dc.identifier.journalClinical Medicine: Arthritis and Musculoskeletal Disordersen
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