✦ LIBER ✦

Homogeneous enzyme immunoassay for macromolecular antigens using hybrid antibody

✍ Scribed by Yoshihiro Ashihara; Isao Nishizono; Hiromasa Suzuki; Yasushi Kasahara

Publisher: John Wiley and Sons
Year: 1987
Tongue: English
Weight: 222 KB
Volume: 1
Category: Article
ISSN: 0887-8013
DOI: 10.1002/jcla.1860010112

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✦ Synopsis

A new homogeneous enzyme immunoassay for the determination of macromolecular antigens has been developed based upon competitive enzyme inhibition using hybrid antibodycontaining anti-ligand and anti-enzyme inhibitory antibodies.

The hybrid antibody was prepared by the reaction of anti-glucose-6-phosphate dehydrogenase (G6PDH) mouse IgG maleimide with anti-human IgM goat IgG-SH. In the absence of the antigen, the hybrid antibody inhibits the enzyme activity. On the other hand, in the presence of an excess amount of antigen, the hybrid antibody binds to the antigen and is unable to bind with the enzyme.

The hybrid antibody inhibited 80% of the original activity of enzyme and saturated the inhibition of G6PDH within 10 min. The enzyme activity was proportional to the concentration of human IgM. The measurable range for IgM was 78 pgiml to 1 mg/ml. Human IgG present in the serum did not affect this method.

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