A rapid method for quantitation of antiviral antibodies.
This paper examines the parameters necessary for the efficient measurement of anti-Theiler's murine encephalomyelitis virus (TMEV) antibodies in an affinity-dependent manner using a variation of a solid-phase particle concentration fluorescence immunoassay (PCFIA). By allowing antibody to react with fluorochrome-labelled virus in fluid phase and subsequently capturing the resulting virus-antibody complexes with anti-immunoglobulin coated polystyrene particles (fluid-phase PCFIA), the present assay allows for both greater sensitivity, specificity and preservation of conformational viral epitopes than do solid-phase immunoassays. Fluid-phase PCFIA proved to be a more rapid quantitative assay than ELISA and significantly diminished non-specific binding by both untreated and heat-inactivated normal mouse sera. This methodology also allowed us to perform competition assays and to determine the dissociation kinetics of anti-viral antibody preparations, investigations which cannot generally be performed as solid-phase immunoassays. Thus fluid-phase PCFIA is a rapid and efficient immunoassay with excellent reproducibility and great versatility.
Peterson
J D
JD
Department of Microbiology-Immunology, Northwestern University Medical School, Chicago, IL 60611.
Kim
J Y
JY
Melvold
R W
RW
Miller
S D
SD
Waltenbaugh
C
C
D000818:Animals; D000914:Antibodies, Viral / Q000032:analysis*; D001666:Binding Sites, Antibody; D001667:Binding, Competitive; D002460:Cell Line; D006224:Cricetinae; D004770:Enterovirus / Q000276:immunology*; D004797:Enzyme-Linked Immunosorbent Assay; D005260:Female; D016650:Fluorescein-5-isothiocyanate; D005452:Fluoresceins; D015200:Fluoroimmunoassay / Q000379:methods* / Q000592:standards; D007700:Kinetics; D004680:Maus Elberfeld virus / Q000276:immunology*; D051379:Mice; D008807:Mice, Inbred BALB C; D008810:Mice, Inbred C57BL; D013861:Thiocyanates
D000914:Antibodies, Viral; D005452:Fluoresceins; D013861:Thiocyanates; D016650:Fluorescein-5-isothiocyanate