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基于使用对放线菌素D有抗性的小鼠骨髓瘤细胞系设计杂交杂交瘤的新方法

[A new approach to the design of hybrid hybridomas based on the use of an actinomycin D-resistant line of murine myeloma].

作者信息

Massino Iu S, Kizim E A, Dmitriev A D

出版信息

Biull Eksp Biol Med. 1991 Nov;112(11):511-4.

PMID:1687361
Abstract

The hybrid hybridomas (tetradomas) were produced from the fusion of the double mutant actinomycin Dr (ADr)/HATs hybridoma to horseradish peroxidase (HRP) and wild type hybridoma to alpha-endorphin (EP). The double mutant phenotype was constructed using the new strategy, based on the fusion of immune mouse splenocytes with mouse myeloma (X63.Ag8, 653) cell variants, made resistant to 30 ng/ml of AD by stepwise selection. This allowed the direct introduction of the dominant selective marker (ADr) into the hybrid cells. Tetradomas secreted the bispecific monoclonal antibodies (bi Mabs), simultaneously binding to EP and HRP in double antigen ELISA, the ELISA plates covered with EP-bovine serum albumin conjugate. Using rat pituitary the bi Mabs were shown to be effective for immunostaining of EP-producing cells. EP-producing cells.

摘要

杂交杂交瘤(四联瘤)由双突变放线菌素D(ADr)/次黄嘌呤氨基蝶呤胸腺嘧啶核苷(HAT)杂交瘤与辣根过氧化物酶(HRP)杂交瘤以及野生型杂交瘤与α-内啡肽(EP)杂交瘤融合产生。双突变表型采用新策略构建,该策略基于免疫小鼠脾细胞与经逐步筛选对30 ng/ml AD产生抗性的小鼠骨髓瘤(X63.Ag8, 653)细胞变体融合。这使得显性选择标记(ADr)能够直接导入杂交细胞。四联瘤分泌双特异性单克隆抗体(双抗),在双抗原酶联免疫吸附测定(ELISA)中,ELISA板包被有EP-牛血清白蛋白偶联物时,双抗能同时结合EP和HRP。利用大鼠垂体,双抗被证明对产生EP的细胞进行免疫染色有效。产生EP的细胞。

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