Rapid biotin-avidin method for quantitation of antiviral antibody isotypes.

Abstract:

: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.

journal_name

J Virol Methods

authors

Peterson JD,Miller SD,Waltenbaugh C

doi

10.1016/0166-0934(90)90135-3

subject

Has Abstract

pub_date

1990-02-01 00:00:00

pages

189-201

issue

2

eissn

0166-0934

issn

1879-0984

pii

0166-0934(90)90135-3

journal_volume

27

pub_type

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