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. 1966 Jan;10(1):81–86.

Assay of anti-hapten antibody with the aid of hapten-coupled bacteriophage

O Mäkelä
PMCID: PMC1423626  PMID: 5930881

Abstract

Treatment of bacteriophage with 3-iodo-4-hydroxy-5-nitrophenyl acetic acid chloride (NIP) in aqueous medium killed a proportion of the phage but the survivors were made susceptible to inactivation by rabbit immune sera to NIP-chicken globulin conjugate. The serum factor inactivating NIP-phage (T2) was eluted from a Sephadex G-200 column as 7S γ-globulin and was neutralized by a sheep antiserum against electrophoretically purified rabbit γ-globulin. The inactivation was strongly inhibited by the hapten and its derivatives. As little as 0.001 micromole per millilitre of NIP—ε-amino caproic acid was inhibitory.

Inactivation of NIP-phage was ascribed to anti-NIP antibody.

Inactivation of NIP-phage (T2) in strong anti-NIP sera approximately followed first-order reaction kinetics until 99 per cent of the phage became inactivated. When incubated with phage for 6 hours concentrations of antiserum as low as 10-7 (1.9×10-5 μg/ml) or less caused measurable inactivation of the phage. The threshold quantity of antibody was estimated to be less than 10-5 μg.

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Selected References

These references are in PubMed. This may not be the complete list of references from this article.

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