PMID- 9266981
OWN - NLM
STAT- MEDLINE
DCOM- 19970908
LR  - 20131121
IS  - 0022-1317 (Print)
IS  - 0022-1317 (Linking)
VI  - 78 ( Pt 8)
DP  - 1997 Aug
TI  - Monoclonal antibodies to the ORF5 product of porcine reproductive and respiratory 
      syndrome virus define linear neutralizing determinants.
PG  - 1867-73
AB  - Complementary DNA encoding the ORF5 gene of a Quebec reference isolate (IAF-Klop) 
      of porcine reproductive and respiratory syndrome virus (PRRSV) was cloned into 
      the prokaryotic expression vectors pGEX-4T and pET21a to produce ORF5-glutathione 
      S-transferase and ORF5-polyhistidine fusion proteins. Five hybridoma cell lines 
      producing monoclonal antibodies (MAbs) to the 25 kDa viral envelope glycoprotein 
      (GP5) were obtained from BALB/c mice immunized with the affinity 
      chromatography-purified GST-ORF5 fusion protein. The polypeptide specificity of 
      these anti-PRRSV MAbs, belonging to the IgG1 isotype, was confirmed by Western 
      immunoblotting assays with recombinant and native viral proteins, and by 
      radioimmunoprecipitation using [35S]methionine-labelled concentrated 
      extracellular virus. All these MAbs showed virus-neutralizing (VN) activity, with 
      VN titres ranging from 1:32 to 1:128. Two MAbs (IAF-1B8 and IAF-8A8) reacted with 
      similar titres with the modified live attenuated vaccine strain ATCC VR-2332, but 
      all five failed to react to the prototype European strain, the Lelystad virus, in 
      VN and indirect immunofluorescence tests. The results obtained suggest that these 
      five anti-PRRSV MAbs are directed to serotype-specific linear neutralizing 
      epitopes which are not affected by the absence of carbohydrate residues.
FAU - Pirzadeh, B
AU  - Pirzadeh B
AD  - Centre de recherche en virologie, Institut Armand-Frappier, Universitedu Quebec, 
      Laval-des-Rapides, Canada.
FAU - Dea, S
AU  - Dea S
LA  - eng
PT  - Journal Article
PT  - Research Support, Non-U.S. Gov't
PL  - England
TA  - J Gen Virol
JT  - The Journal of general virology
JID - 0077340
RN  - 0 (Antibodies, Monoclonal)
RN  - 0 (Antigens, Viral)
RN  - 0 (Immunoglobulin G)
RN  - 0 (Peptides)
RN  - 0 (Recombinant Fusion Proteins)
RN  - 26062-48-6 (polyhistidine)
RN  - 4QD397987E (Histidine)
RN  - EC 2.5.1.18 (Glutathione Transferase)
SB  - IM
MH  - Animals
MH  - *Antibodies, Monoclonal
MH  - Antigens, Viral/biosynthesis/chemistry/*immunology
MH  - Blotting, Western
MH  - Cloning, Molecular
MH  - Glutathione Transferase
MH  - *Histidine
MH  - Immunoglobulin G
MH  - Mice
MH  - Mice, Inbred BALB C
MH  - Neutralization Tests
MH  - Peptides
MH  - Porcine respiratory and reproductive syndrome virus/genetics/*immunology
MH  - Radioimmunoassay
MH  - Recombinant Fusion Proteins/biosynthesis/chemistry/immunology
EDAT- 1997/08/01 00:00
MHDA- 1997/08/01 00:01
CRDT- 1997/08/01 00:00
PHST- 1997/08/01 00:00 [pubmed]
PHST- 1997/08/01 00:01 [medline]
PHST- 1997/08/01 00:00 [entrez]
AID - 10.1099/0022-1317-78-8-1867 [doi]
PST - ppublish
SO  - J Gen Virol. 1997 Aug;78 ( Pt 8):1867-73. doi: 10.1099/0022-1317-78-8-1867.