NDLI: High-resolution epitope mapping for monoclonal antibodies to the structural protein Erns of classical swine fever virus using peptide array and random peptide phage display approaches.

Content Provider	World Health Organization (WHO)-Global Index Medicus
Author	Lin, Min McRae, Heather Dan, Hanhong Tangorra, Erin Laverdiere, Amanda Pasick, John
Description	Country affiliation: Canada Author Affiliation: Lin M ( Canadian Food Inspection Agency, Ottawa Laboratory (Fallowfield), Ottawa, ON, Canada. linm@inspection.gc.ca)
Abstract	The structural glycoprotein E(rns) (an envelope protein with RNase activity) of classical swine fever virus (CSFV) is not well characterized with respect to its antigenic structure and organization. Here, we investigated the antigenic sites on E(rns) by raising mAbs against the Escherichia coli expressed E(rns) of CSFV strain Alfort/187 and defined the B-cell epitopes recognized by these antibodies. Eighteen mAbs to E(rns) were identified and they were classified as either immunoglobulin subclass G1 or G2b. Using an array of overlapping 12-mer peptides, spanning aa 27-227 of E(rns), the epitopes for 12 mAbs were mapped to a high resolution of six to eight residues, which cluster in five discrete locations, ¹³GIWPEKIC³8 (group I), 65NYTCCKLQ7² (group II), ¹²7QARNRPTT¹³4 (group III), ¹45SFAGTVIE¹5² (group IV) and ¹6¹VEDILY¹66 (group V). Two mAbs recognize two or more antigenic determinants, including the group II epitope. The epitopes for four other mAbs could not be mapped using the overlapping 12-mer peptides. Random peptide phage display with one mAb from each of all the groups except group V further identified some conserved residues that may be critical for binding antibodies, i.e. Trp³³ in the epitope of group I, Leu7¹ in the epitope of group II, Gln¹²7 and Apn¹³° in the epitope of group III, and Ser¹45 and Gly¹48 in the epitope of group IV. This study has provided new insights into the structure and organization of epitopes on the CSFV E(rns) and valuable epitope information for the rational design of vaccines, drugs and diagnostic immunoassays for CSFV.
File Format	HTM / HTML
ISSN	00221317
Issue Number	Pt 12
Volume Number	91
e-ISSN	14652099
Journal	Journal of General Virology
Language	English
Publisher	Microbiology Society
Publisher Date	2010-12-01
Publisher Place	Great Britain (UK)
Access Restriction	Subscribed
Subject Keyword	Epitope Mapping Discipline Virology Immunoglobulin G Isolation & Purification Mice, Inbred Balb C Ribonucleases Antibodies, Monoclonal Peptide Library Antibodies, Viral Epitopes, B-lymphocyte Journal Article Viral Structural Proteins Immunology Animals Conserved Sequence Protein Array Analysis Female Mice Classical Swine Fever Virus Methods
Content Type	Text
Resource Type	Article
Subject	Virology

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