A method for rapid, ligation-independent reformatting of recombinant monoclonal antibodies
A method for rapid, ligation-independent reformatting of recombinant monoclonal antibodies
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DOI：
10.1016/j.jim.2010.02.001
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发表时间：
2010-03-31
影响因子：
2.2
通讯作者：
Munro, Trent P.
中科院分区：
文献类型：
--
作者：
Jones, Martina L.;Seldone, Therese;Munro, Trent P.
重组单克隆抗体目前主导蛋白质生物制剂市场。从靶抗原发现和筛选到重组治疗性抗体的路径可能是耗时且费力的。我们描述了一组表达载体，称为mAbXpress，能够快速和序列无关的抗体可变区插入到人恒定区骨架。该方法利用Clontech的In Fusion（TM）克隆系统，其允许可变区盒的无连接、高效插入而不添加外源氨基酸。这些模块化载体简化了在治疗或诊断候选物的初步评估期间的抗体重新格式化过程。所得构建体可直接用于哺乳动物细胞中的瞬时或可扩增的稳定表达。该方法的有效性通过产生功能性的完全人源抗人CD83单克隆抗体来证明。皇冠版权所有（C）2010由爱思唯尔B.V.出版保留所有权利。
Recombinant monoclonal antibodies currently dominate the protein biologics marketplace. The path from target antigen discovery and screening, to a recombinant therapeutic antibody can be time-consuming and laborious. We describe a set of expression vectors, termed mAbXpress, that enable rapid and sequence-independent insertion of antibody variable regions into human constant region backbones. This method takes advantage of the In Fusion (TM) cloning system from Clontech, which allows ligation-free, high-efficiency insertion of the variable region cassette without the addition of extraneous amino acids. These modular vectors simplify the antibody reformatting process during the preliminary evaluation of therapeutic or diagnostic candidates. The resulting constructs can be used directly for transient or amplifiable, stable expression in mammalian cells. The effectiveness of this method was demonstrated by the creation of a functional, fully human anti-human CD83 monoclonal antibody. Crown Copyright (C) 2010 Published by Elsevier B.V. All rights reserved.