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.
De Lau
W B
WB
Department of Clinical Immunology, University Hospital, Utrecht, The Netherlands.
Van Loon
A E
AE
Heije
K
K
Valerio
D
D
Bast
B J
BJ
D000630:Aminopterin; D000918:Antibody Specificity; D001375:Azaguanine; D002459:Cell Fusion; D002460:Cell Line; D004351:Drug Resistance; D006801:Humans; D006825:Hybridomas / Q000032:analysis / Q000378:metabolism*; D019271:Hypoxanthine; D007042:Hypoxanthines / Q000276:immunology; D007958:Leukocyte Count; D009154:Mutation; D009355:Neomycin; D010641:Phenotype; D013936:Thymidine; D014161:Transduction, Genetic
D007042:Hypoxanthines; D019271:Hypoxanthine; D009355:Neomycin; D000630:Aminopterin; D001375:Azaguanine; D013936:Thymidine