PMID: 46693Feb 1, 1975

Preparation and standardization of perotidase-labeled anti-human IgG antibody for use in determination of serum antinuclear antibody levels

American Journal of Clinical Pathology
E R WalwickR M Nakamura

Abstract

Peroxidase-labeled antibody conjugates were prepared by a two-step conjugation procedure using glutaraldehyde. Immunoadsorbent-purified antibody and the gamma-globulin fraction of sheep anti-human IgG antiserum were employed for these preparations, Procedures for the determination of the antibody and enzymatic activity, as well as the specificity of the enzy-e-antibody reactions, were outlined. Peroxidase-anti-human-IgG conjugates were prepared with approximately a 1:1 molar ratio of peroxidase to IgG, which demonstrated a loss of 10-15% of precipitating antibody activity. Standardization procedures for use of peroxidase-labeled antibody in the indirect ANA test were established. The peroxidase-labeled antibody in the indirect ANA test were established. The peroxidase-antibody procedure was found to demonstrate a reproducible plateau endpoint which was comparable to that of the fluorescent antibody preparations.

Related Concepts

Ovoperoxidase
Immunofluorescence Assay
Conjugation
Peroxidases
Immune Sera
Dall Sheep
Fluorescent Antinuclear Antibodies
Gamma globulin
Sheep antigen
Molar Tooth

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