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Clinical Chemistry 34: 1693-1696, 1988;
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Clinical Chemistry, Vol 34, 1693-1696, Copyright © 1988 by American Association for Clinical Chemistry

Production of murine hybrid-hybridomas secreting bispecific monoclonal antibodies for use in urease-based immunoassays

M Takahashi and SA Fuller
Allelix Diagnostics, Inc., Mississauga, Ontario, Canada.

To produce bispecific antibodies (BiAbs) for enzyme immunoassay (EIA) to replace antibody-enzyme conjugates, we developed a panel of 8- azaguanine/ouabain-resistant anti-urease variant hybridoma cell lines for use in hybridoma-hybridoma fusions. These variants represent mouse immunoglobulin subclasses IgM, IgG1, IgG2a, and IgG2b and have growth rates equal to those of the parental hybridomas. We fused an anti- urease-secreting variant hybridoma with an anti-human choriogonadotropin (hCG)-secreting hybridoma (both of IgG1 subclass) and selected the desired product with growth media containing hypoxanthine-aminopterin-thymidine (HAT) and ouabain. Over 95% of the resulting hybrids secreted anti-urease, and 60% of these secreted anti- hCG. The bispecific nature of secreted antibodies was demonstrated in a simultaneous EIA where BiAbs, hCG, and urease (EC 3.5.1.5) were incubated together in anti-hCG-coated microwells. As little as 25 int. units of hCG per liter could be reliably detected, which is equivalent to that for antibody-enzyme conjugates in EIA.


The following articles in journals at HighWire Press have cited this article:


Home page
 Clin. Chem.Home page
K. Morimoto and K. Inouye
Application of Bispecific F(ab')2{micro} Fragments Prepared from IgMs against Carcinoembryonic Antigen and Alkaline Phosphatase
Clin. Chem., September 1, 2000; 46(9): 1492 - 1493.
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Copyright © 1988 by the American Association for Clinical Chemistry.