Rapid biotin-avidin method for quantitation of antiviral antibody isotypes.
A rapid and efficient method is described for isotype quantitation of antiviral antibodies in mice infected with Theiler's murine encephalomyelitis virus (TMEV). Serum antibodies were reacted with fluorochrome-labeled TMEV in a modified fluid-phase particle concentration fluorescence immunoassay (PCFIA). Biotin and avidin were used to attach anti-immunoglobulin isotype antibodies to polystyrene particles by the separate incubation of biotinylated goat anti-mouse isotypes (IgG1-, IgG2a-, IgG2b-, IgG3-, or IgM-specific) with avidin coupled polystyrene particles. These anti-isotype particles captured the virus-antibody complexes. Mouse myeloma proteins were used to quantitate and standardize isotype profiles of normal mouse serum using fluorescein isothiocyanate (FITC)-labeled, goat anti-mouse isotypes and polystyrene particles coated with goat anti-mouse. These assays quantitated the affinity-purified mouse serum antiviral antibodies for the standardization of antiviral isotype assays. Immunoglobulin of all serum isotypes as well as the amount of virus-specific isotypes can be quantitated rapidly and accurately.
Peterson
J D
JD
Department of Microbiology-Immunology, Northwestern University Medical School, Chicago, IL 60611.
Miller
S D
SD
Waltenbaugh
C
C
D000818:Animals; D000914:Antibodies, Viral / Q000032:analysis* / Q000592:standards; D001360:Avidin; D001710:Biotin; D004770:Enterovirus / Q000276:immunology*; D004769:Enterovirus Infections / Q000276:immunology; D005260:Female; D007132:Immunoglobulin Isotypes / Q000032:analysis* / Q000592:standards; D004680:Maus Elberfeld virus / Q000276:immunology*; D051379:Mice; D014773:Virology / Q000379:methods
D000914:Antibodies, Viral; D007132:Immunoglobulin Isotypes; D001360:Avidin; D001710:Biotin