Epitopic mapping of linear and conformation-dependent antigenic determinants on GP5 of five U.S. bluetongue viruses

Yi Yuan Yang, Todd M. Johnson, James 0. Mecham, James P. Tam, Joseph K K Li

Research output: Contribution to journalArticle

14 Citations (Scopus)

Abstract

Two distinct antigenic determinants of the major outer capsid protein, GP5, of five U.S. bluetongue viruses have been identified and mapped using monoclonal and oligoclonal antibodies. One antigenic site, identified by oligoclonal antibody AK-15, was found to be common and conserved in all five U.S. BTV serotypes. This linear epitope was located between amino acid residues 175 and 189 (ALQREAAERSEDEIK). The second determinant identified by monoclonal antibody 34.7 was present in BTV-2, -10, -11, and -17 but absent in BTV-13. The binding of this monoclonal antibody to GP5 could be blocked specifically by one of three short synthetic peptides located among amino acid residues 33-42 (KAAERFAESE), 159-168 (EKILKEEDSK), and 206-215 (EIERDGMQEE), indicating that this antigenic determinant is conformation-dependent. Oligoclonal antibody (AK-15) reacted with denatured GP5 immobilized on nitrocellulose membrane after Western transfer as well as with native GP5 present on the surface of purified BTV virions. Monoclonal antibody (34.7) reacted only with denatured GP5 but not native GP5 using an ELISA assay. However, these two antigenic epitopes alone did not elicit detectable neutralizing antibodies as determined by plaque reduction assay.

Original languageEnglish
Pages (from-to)530-536
Number of pages7
JournalVirology
Volume188
Issue number2
DOIs
Publication statusPublished - 1992
Externally publishedYes

Fingerprint

Bluetongue virus
Epitopes
Monoclonal Antibodies
Amino Acids
Collodion
Antibodies
Capsid Proteins
Neutralizing Antibodies
Virion
Enzyme-Linked Immunosorbent Assay
Peptides
Membranes
difluprednate

ASJC Scopus subject areas

  • Virology
  • Infectious Diseases

Cite this

Epitopic mapping of linear and conformation-dependent antigenic determinants on GP5 of five U.S. bluetongue viruses. / Yang, Yi Yuan; Johnson, Todd M.; Mecham, James 0.; Tam, James P.; Li, Joseph K K.

In: Virology, Vol. 188, No. 2, 1992, p. 530-536.

Research output: Contribution to journalArticle

Yang, Yi Yuan ; Johnson, Todd M. ; Mecham, James 0. ; Tam, James P. ; Li, Joseph K K. / Epitopic mapping of linear and conformation-dependent antigenic determinants on GP5 of five U.S. bluetongue viruses. In: Virology. 1992 ; Vol. 188, No. 2. pp. 530-536.
@article{c79a2e44e5b34076b3a48721375cf450,
title = "Epitopic mapping of linear and conformation-dependent antigenic determinants on GP5 of five U.S. bluetongue viruses",
abstract = "Two distinct antigenic determinants of the major outer capsid protein, GP5, of five U.S. bluetongue viruses have been identified and mapped using monoclonal and oligoclonal antibodies. One antigenic site, identified by oligoclonal antibody AK-15, was found to be common and conserved in all five U.S. BTV serotypes. This linear epitope was located between amino acid residues 175 and 189 (ALQREAAERSEDEIK). The second determinant identified by monoclonal antibody 34.7 was present in BTV-2, -10, -11, and -17 but absent in BTV-13. The binding of this monoclonal antibody to GP5 could be blocked specifically by one of three short synthetic peptides located among amino acid residues 33-42 (KAAERFAESE), 159-168 (EKILKEEDSK), and 206-215 (EIERDGMQEE), indicating that this antigenic determinant is conformation-dependent. Oligoclonal antibody (AK-15) reacted with denatured GP5 immobilized on nitrocellulose membrane after Western transfer as well as with native GP5 present on the surface of purified BTV virions. Monoclonal antibody (34.7) reacted only with denatured GP5 but not native GP5 using an ELISA assay. However, these two antigenic epitopes alone did not elicit detectable neutralizing antibodies as determined by plaque reduction assay.",
author = "Yang, {Yi Yuan} and Johnson, {Todd M.} and Mecham, {James 0.} and Tam, {James P.} and Li, {Joseph K K}",
year = "1992",
doi = "10.1016/0042-6822(92)90507-L",
language = "English",
volume = "188",
pages = "530--536",
journal = "Virology",
issn = "0042-6822",
publisher = "Academic Press Inc.",
number = "2",

}

TY - JOUR

T1 - Epitopic mapping of linear and conformation-dependent antigenic determinants on GP5 of five U.S. bluetongue viruses

AU - Yang, Yi Yuan

AU - Johnson, Todd M.

AU - Mecham, James 0.

AU - Tam, James P.

AU - Li, Joseph K K

PY - 1992

Y1 - 1992

N2 - Two distinct antigenic determinants of the major outer capsid protein, GP5, of five U.S. bluetongue viruses have been identified and mapped using monoclonal and oligoclonal antibodies. One antigenic site, identified by oligoclonal antibody AK-15, was found to be common and conserved in all five U.S. BTV serotypes. This linear epitope was located between amino acid residues 175 and 189 (ALQREAAERSEDEIK). The second determinant identified by monoclonal antibody 34.7 was present in BTV-2, -10, -11, and -17 but absent in BTV-13. The binding of this monoclonal antibody to GP5 could be blocked specifically by one of three short synthetic peptides located among amino acid residues 33-42 (KAAERFAESE), 159-168 (EKILKEEDSK), and 206-215 (EIERDGMQEE), indicating that this antigenic determinant is conformation-dependent. Oligoclonal antibody (AK-15) reacted with denatured GP5 immobilized on nitrocellulose membrane after Western transfer as well as with native GP5 present on the surface of purified BTV virions. Monoclonal antibody (34.7) reacted only with denatured GP5 but not native GP5 using an ELISA assay. However, these two antigenic epitopes alone did not elicit detectable neutralizing antibodies as determined by plaque reduction assay.

AB - Two distinct antigenic determinants of the major outer capsid protein, GP5, of five U.S. bluetongue viruses have been identified and mapped using monoclonal and oligoclonal antibodies. One antigenic site, identified by oligoclonal antibody AK-15, was found to be common and conserved in all five U.S. BTV serotypes. This linear epitope was located between amino acid residues 175 and 189 (ALQREAAERSEDEIK). The second determinant identified by monoclonal antibody 34.7 was present in BTV-2, -10, -11, and -17 but absent in BTV-13. The binding of this monoclonal antibody to GP5 could be blocked specifically by one of three short synthetic peptides located among amino acid residues 33-42 (KAAERFAESE), 159-168 (EKILKEEDSK), and 206-215 (EIERDGMQEE), indicating that this antigenic determinant is conformation-dependent. Oligoclonal antibody (AK-15) reacted with denatured GP5 immobilized on nitrocellulose membrane after Western transfer as well as with native GP5 present on the surface of purified BTV virions. Monoclonal antibody (34.7) reacted only with denatured GP5 but not native GP5 using an ELISA assay. However, these two antigenic epitopes alone did not elicit detectable neutralizing antibodies as determined by plaque reduction assay.

UR - http://www.scopus.com/inward/record.url?scp=0026655101&partnerID=8YFLogxK

UR - http://www.scopus.com/inward/citedby.url?scp=0026655101&partnerID=8YFLogxK

U2 - 10.1016/0042-6822(92)90507-L

DO - 10.1016/0042-6822(92)90507-L

M3 - Article

C2 - 1374982

AN - SCOPUS:0026655101

VL - 188

SP - 530

EP - 536

JO - Virology

JF - Virology

SN - 0042-6822

IS - 2

ER -