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从对CD3和卵巢癌具有特异性的小鼠三瘤OC/TR中纯化双特异性F(ab')2 。

Purification of bispecific F(ab')2 from murine trinoma OC/TR with specificity for CD3 and ovarian cancer.

作者信息

Warnaar S O, De Paus V, Lardenoije R, Machielse B N, De Graaf J, Bregonje M, Van Haarlem H

机构信息

Centocor B.V., Leiden, The Netherlands.

出版信息

Hybridoma. 1994 Dec;13(6):519-26. doi: 10.1089/hyb.1994.13.519.

Abstract

A study was made of the stability of the murine bispecific trinoma OC/TR with respect to secretion of both types of parental heavy and light chains. OC/TR is a cell line producing bispecific antibody that reacts with the CD3 antigen on T cells and the folate-binding receptor--frequently found to be overexpressed on ovarian carcinoma cells. Of the 10 different IgG combinations theoretically possible with 2 heavy and 2 light chains, 6 combinations were secreted. Subclones varied considerably in relative production of the two parental heavy and light chains. A detailed analysis was made of the binding characteristics and retargeting activity of each of the IgGs produced. From a clone producing a relatively high quantity of bispecific IgG, a large-scale production was initiated. The purification of clinical grade bispecific F(ab')2 from harvest fluids is described. The yield from this purification process was found to be comparable to the yield of bispecific F(ab')2 after chemical cross-linking of two different Fab'.

摘要

对鼠双特异性肿瘤细胞系OC/TR在两种亲本重链和轻链分泌方面的稳定性进行了研究。OC/TR是一种产生双特异性抗体的细胞系,该抗体与T细胞上的CD3抗原以及叶酸结合受体发生反应,而叶酸结合受体在卵巢癌细胞上经常过度表达。在理论上由2条重链和2条轻链组成的10种不同IgG组合中,有6种组合被分泌。亚克隆在两种亲本重链和轻链的相对产量上有很大差异。对所产生的每种IgG的结合特性和重定向活性进行了详细分析。从产生相对大量双特异性IgG的克隆开始进行大规模生产。描述了从收获液中纯化临床级双特异性F(ab')2的方法。发现该纯化过程的产量与两种不同Fab'化学交联后双特异性F(ab')2的产量相当。

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