De Lau W B, Van Loon A E, Heije K, Valerio D, Bast B J
Department of Clinical Immunology, University Hospital, Utrecht, The Netherlands.
J Immunol Methods. 1989 Feb 8;117(1):1-8. doi: 10.1016/0022-1759(89)90111-7.
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.
本文描述了一种制备产生双特异性抗体的杂交杂交瘤的详细方法。这是通过融合两个表型不同的杂交瘤细胞系（HAT(s)/neo(r) 和 HAT(r)/neo(s)）来实现的，从而能够选择合适的杂交细胞。通过8-氮杂鸟嘌呤处理从两个融合亲本之一获得HATs突变体；这些突变表型的出现频率出乎意料地高。为了在其中一个HAT(s)融合亲本中引入显性neo(r)标记基因，使用了逆转录病毒载体以获得高效的基因转移。我们的方法在产生所谓的四瘤杂交杂交瘤方面非常有效。双特异性抗体的检测基于一种抗体同时结合两种不同抗原，或者基于该分子中存在两种不同的重链同种型。
