Chemistry: molecular biology and microbiology – Measuring or testing process involving enzymes or... – Involving antigen-antibody binding – specific binding protein...
Patent
1979-01-08
1981-03-17
Wiseman, Thomas G.
Chemistry: molecular biology and microbiology
Measuring or testing process involving enzymes or...
Involving antigen-antibody binding, specific binding protein...
435177, 435188, 435192, 23230B, 424 12, C12Q 166, C12N 1102, C12N 996
Patent
active
042568331
ABSTRACT:
Horse radish peroxidase (HRP) is treated with phenyl isothiocyanate (PITC) to block the free amino groups on the enzyme. The PITC derivative of HRP is treated with periodate to oxidize the carbohydrate moiety on the enzyme, thus generating aldehyde groups. Gamma G globulin fraction (IgG) purified from an anti-human IgE serum is conjugated to the peroxidase-aldehyde by formation of a Schiff's base between the aldehyde group on the enzyme and the amino groups on the IgG. The Schiff's base is stabilized by reduction using the optimal amounts of sodium borohydride determined by tiration. A stable HRP-anti IgE IgG conjugate prepared thus is employed in a solid phase enzyme immunoassay for the detection of allergen specific IgE. The results of this assay can be used to determine a safe initial hypersensitization dosage level.
REFERENCES:
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patent: 4031197 (1977-06-01), Marinkovich
patent: 4069105 (1978-01-01), Singh
patent: 4115539 (1978-09-01), Eisenhardt et al.
Berezin et al., "Catalytic Properties and Thermostability of Horseradish Peroxidase Covalently Bound to Sepharose Through Carbohydrate Residues of the Enzyme", Chem. Abstracts, vol. 87, No. 23, p. 165 (1977) abs. #35084d.
Mukajima, et al., "Enzyme Immunoassay Using Multiple Antigen Enzyme Complex", Chem. Abstracts, vol. 87, No. 23, p. 269 (1977) abs. #180244K.
March, Advanced Organic Chemistry: Reactions, Mechanisms, and Structures, McGraw-Hill Book Co., New York (1968) pp. 890, 891.
Ali Majid
Fayemi Alfred
Mesa-Tejada Ricardo
Nalebuft Donald
Ramanarayanan Madhava P.
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