二价(发动机)
分子生物学
克隆(编程)
抗体
分子克隆
噬菌体展示
表达式向量
计算生物学
生物
抗原
化学
基因
遗传学
重组DNA
基因表达
计算机科学
有机化学
金属
程序设计语言
作者
Rico Ballmann,Kai-Thomas Schneider,Kristian Daniel Ralph Roth,Stefan Dübel
出处
期刊:Methods in molecular biology
日期:2023-01-01
卷期号:: 411-417
标识
DOI:10.1007/978-1-0716-3381-6_21
摘要
The antigen-binding ability of each antibody clone selected by phage display is usually initially ranked by a screening ELISA using monovalent scFv antibody fragments. Further characterization often requires bivalent antibody molecules such as IgG or scFv-Fc fusions. To produce these, the V region encoding genes of selected hits have to be cloned into a mammalian expression vector and analyzed as a bivalent molecule, requiring a laborious cloning procedure. We established a high-throughput procedure allowing rapid screening of candidates in bivalent formats. This protocol allows for the parallelized cloning of all selected antibody fragments into a mammalian expression vector in the 96-well plate format. The bivalent antibody molecules can then be produced and purified in 96-well plates for further analysis in microtiter plate assays.
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