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PubMed1989

Production of hybrid hybridomas based on HAT(s)-neomycin(r) double mutants.

A detailed procedure is described for the preparation of hybrid hybridomas, that produce bispecific antibodies. This is achieved by fusing two hybridoma cell lines that are phenotypically distinct (HAT(s)/neo(r) and HAT(r)/neo(s)) and thereby allow for the selection of the appropriate hybrid cells. HATs mutants were obtained from one of the two fusion partners by 8-azaguanine treatment; these mutant phenotypes were found in an unexpected high frequency. For the introduction of the dominant neo(r) marker gene in one of the HAT(s) fusion partners, a retroviral vector was used in order to obtain a high efficiency of gene transfer. Our method was very effective in the production of hybrid hybridomas, so-called quadromas. The detection of bispecific antibodies was based on simultaneous binding by one antibody of two different antigens, or on the presence of two different H chain isotypes in this molecule.

Aminopterin↗