Purification of infectious canine parvovirus from cell culture by affinity chromatography with monoclonal antibodies.
Immuno affinity chromatography with virus neutralizing monoclonal antibodies, directed to the haemagglutinating protein of canine parvovirus (CPV) was used to purify and concentrate CPV from infected cell culture. The procedure was monitored by testing the respective fractions in an infectivity titration system, in an ELISA, in a haemagglutination assay and by negative contrast electron microscopy to quantify CPV or CPV antigen. The degree of purification was further estimated by testing the fractions for total protein content in a colorimetric method, for bovine serum albumin content in an ELISA and by SDS-PAGE. Over 99% of the contaminating proteins proved to be removed, and 20% or 70-90% of infectious CPV or CPV antigen, respectively, was recovered.
Rimmelzwaan
G F
GF
Groen
J
J
Juntti
N
N
Teppema
J S
JS
UytdeHaag
F G
FG
Osterhaus
A D
AD
D000818:Animals; D000911:Antibodies, Monoclonal / Q000276:immunology*; D000914:Antibodies, Viral / Q000276:immunology*; D002460:Cell Line; D002846:Chromatography, Affinity / Q000379:methods*; D004285:Dogs; D006389:Hemagglutinins, Viral / Q000276:immunology; D007158:Immunologic Techniques; D010321:Parvoviridae / Q000276:immunology / Q000302:isolation & purification*; D014776:Virus Cultivation
D000911:Antibodies, Monoclonal; D000914:Antibodies, Viral; D006389:Hemagglutinins, Viral