Production of a single chain anti-CEA antibody from the hybridoma cell line T84.66 using a modified colony-lift selection procedure to detect antigen-positive ScFv bacterial clones.
Production of a single chain anti-CEA antibody from the hybridoma cell line T84.66 using a modified colony-lift selection procedure to detect antigen-positive ScFv bacterial clones.
复制标题
使用改良的集落提升选择程序从杂交瘤细胞系 T84.66 生产单链抗 CEA 抗体，以检测抗原阳性 ScFv 细菌克隆。
DOI：
10.1089/hyb.1998.17.1
复制
发表时间：
1998
期刊：
影响因子：
--
通讯作者：
Khazaeli,MB
中科院分区：
文献类型：
--
作者：
Rodenburg,CM;Mernaugh,R;Bilbao,G;Khazaeli,MB
与小鼠单克隆抗体 (MAb) 相比，重组单链 Fv (scFv) 抗体具有许多优势，例如更快地从血液中清除、改善肿瘤定位、减少人抗小鼠抗体 (HAMA) 反应以及可通过遗传方法操纵 scFv。 scFv 抗体（指定为 RK10.2）是使用抗 CEA T84.66 杂交瘤细胞作为遗传起始材料来源和法玛西亚重组噬菌体抗体系统 (RPAS) 生成的。使用改良的菌落寿命选择程序和抗原包被的过滤器鉴定表达抗原阳性可溶性 scFv 的大肠杆菌。所得抗CEA scFv（命名为RK10.2）的分子量约为33.6 kDa，15℃时等电点为5.2。 RK10.2 scFv 与带有 CEA 抗原的 LS174 T 细胞相互作用，并抑制 ELISA 中的抗 CEA MAb/CEA 抗原相互作用以及 RIA 中的抗 CEA MAb/LS174 T 细胞相互作用。改进的集落提升方法避开了通常用于亲和选择抗原阳性 scFv 克隆的更耗时的噬菌体展示方法。
Recombinant single chain Fv (scFv) antibodies offer many advantages over mouse monoclonal antibodies (MAbs) such as faster clearance from blood, improved tumor localization, reduced human anti-mouse antibody (HAMA) response, and the availability to manipulate the scFv through genetic approaches. The scFv antibody (designated RK10.2) was generated using anti-CEA T84.66 hybridoma cells as a source of genetic starting material and the Pharmacia Recombinant Phage Antibody System (RPAS).Escherichia coliclones expressing antigen-positive soluble scFv were identified using a modified colony-life selection procedure and antigen-coated filters. The resultant anti-CEA scFv (designated RK10.2) had a molecular weight of approximately 33.6 kDa and an isoelectric point of 5.2 at 15°C. The RK10.2 scFv interacted with LS174 T cells bearing the CEA antigen and inhibited the anti-CEA MAb/CEA antigen interaction in ELISA and the anti-CEA MAb/LS174 T cell interaction in a RIA. The modified colony-lift approach circumvented the more time-consuming phage-display approach that is normally taken to affinity select for antigen-positive scFv clones.