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Hoffmann, R. A wiki for the life sciences where authorship matters. Nature Genetics (2008)

Production of hybrids secreting bispecific antibodies recognising CEA and doxorubicin.

A monoclonal anti-CEA secreting hybridoma (11-285-14) was made hypoxanthine, aminopterin and thymidine (HAT) sensitive by back selecting it in increasing concentrations of 8-azaguanine. Eight 8-azaguanine resistant fusion partners were selected based on growth characteristics and continued anti-CEA production. Since doxorubicin (Dox) is a hapten, it was conjugated to the carrier proteins keyhole limpet hemocyanin (KLH) or bovine serum albumin ( BSA) using 1-ethyl-3- (dimethyl-aminopropyl) carbodiimide. Dox-KLH and Dox- BSA conjugates were used to immunize mice and spleen cells from these mice were used for fusions with the HAT sensitive anti-CEA partner using standard hybridoma procedures. Enzyme linked immunosorbent assays (ELISAs) were developed to test the hybrids obtained for anti-CEA, anti-DOX, anti- BSA and bispecific monoclonal antibody (BsMab) activity. Sixteen fusions with spleen cells from DOX-KLH immunized mice yielded 621 hybrids of which 47 showed low level BsMab activity by ELISA. Eight fusions with spleen cells from DOX- BSA immunized mice yielded 297 hybrids. Fifty of these hybrids showing dual reactivity have been cloned and subcloned to yield 7 subclones with stable BsMab activity for CEA and doxorubicin.[1]


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