Immunoaffinity purification of baculovirus-expressed rubella virus E1 for diagnostic purposes.
Three monoclonal antibodies, termed 4E10, 1E11:10, and 2D9:1, were generated against rubella virus. Immunoblot analysis with purified authentic rubella virus or recombinant baculovirus-expressed rubella virus structural proteins E1, E2, and C demonstrated that they were directed against the E1 envelope glycoprotein of the rubella virus particle. By using the yeast Ty virus-like particle system, it was possible to map the binding site of 1E11:10 within amino acids 236 to 286 of the E1 protein and the binding sites of 2D9:1 and 4E10 outside this region. Immunoaffinity purification with these monoclonal antibodies made it evident that they are useful for obtaining large quantities of pure baculovirus-expressed rubella virus envelope protein E1. The diagnostic potential of this immunoaffinity-purified recombinant rubella virus E1 protein compared with that of authentic rubella virus is demonstrated.
Lindqvist
C
C
Department of Biochemistry and Pharmacy, Abo Akademi University, Turku, Finland.
Schmidt
M
M
Heinola
J
J
Jaatinen
R
R
Osterblad
M
M
Salmi
A
A
Keränen
S
S
Akerman
K
K
Oker-Blom
C
C
D000818:Animals; D000911:Antibodies, Monoclonal / Q000276:immunology*; D000914:Antibodies, Viral / Q000276:immunology*; D006801:Humans; D007163:Immunosorbent Techniques; D051379:Mice; D008807:Mice, Inbred BALB C; D008863:Microspheres; D009036:Moths; D017924:Nucleopolyhedroviruses; D011993:Recombinant Fusion Proteins / Q000096:biosynthesis / Q000276:immunology / Q000302:isolation & purification*; D012409:Rubella / Q000097:blood* / Q000276:immunology; D012412:Rubella virus / Q000235:genetics / Q000276:immunology*; D012441:Saccharomyces cerevisiae; D014759:Viral Envelope Proteins / Q000096:biosynthesis / Q000276:immunology / Q000302:isolation & purification*
D000911:Antibodies, Monoclonal; D000914:Antibodies, Viral; D011993:Recombinant Fusion Proteins; D014759:Viral Envelope Proteins; C040901:rubella antibodies; C061826:E1 envelope protein, Rubella virus