Isolation and recombinant expression of an MHV-JHM neutralising monoclonal antibody.
The monoclonal antibody A1 (mab A1) efficiently neutralises the infection of susceptible cells by the murine hepatitis virus MHV-JHM in vitro and in vivo (Wege et al., 1984). The variable regions of mab A1 were amplified from mRNA of the respective hybridoma cell line by RT-PCR and integrated into different eukaryotic expression vectors. The biological function of the recombinant antibody constructs was verified by virus neutralisation assays. Whereas a complete recombinant antibody (mab A1rec.) expressed in transfected murine myeloma cells inhibited the MHV-JHM infection as well as the parental antibody, a single-chain Fv derived from mab A1 did not show any neutralising activity.
Institute of Virology and Immunology, University of Würzburg, Germany.
A F
AF
Kolb
M
M
Lechermaier
A
A
Heister
A
A
Toksoy
S G
SG
Siddell
D000818:Animals; D000911:Antibodies, Monoclonal / Q000096:biosynthesis* / Q000276:immunology / Q000302:isolation & purification; D000914:Antibodies, Viral / Q000096:biosynthesis* / Q000276:immunology / Q000302:isolation & purification; D002460:Cell Line; D007135:Immunoglobulin Variable Region / Q000096:biosynthesis* / Q000276:immunology / Q000302:isolation & purification; D051379:Mice; D006517:Murine hepatitis virus / Q000276:immunology*; D009500:Neutralization Tests; D011993:Recombinant Fusion Proteins / Q000096:biosynthesis; D014407:Tumor Cells, Cultured
D000911:Antibodies, Monoclonal; D000914:Antibodies, Viral; D007135:Immunoglobulin Variable Region; D011993:Recombinant Fusion Proteins