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Single-step colony assay with autoinduction of scFv expression for the screening of antibody libraries.
Biotechniques ( IF 2.7 ) Pub Date : 2019-02-20 , DOI: 10.2144/btn-2018-0195
Mieko Kato 1 , Yoshiro Hanyu 2
Affiliation  

We present a simple colony assay method for screening antibody libraries based on autoinduction of antibody fragment expression. This protocol eliminates the need for colony size monitoring and a separate induction step for single-chain Fv (scFv) antibody fragment expression. Here, scFvs are expressed in an automatic and timely fashion during the assay, resulting in high yields of positive clones and substantial time savings. The method was used successfully to establish monoclonal scFvs with high affinity and specificity against human IgG.

中文翻译:

自动筛选scFv表达的单步菌落测定法用于抗体库的筛选。

我们提出了一种基于自动诱导的抗体片段表达筛选抗体库的简单菌落分析方法。该协议消除了对菌落大小监控和单链Fv(scFv)抗体片段表达的单独诱导步骤的需要。在此,scFvs在检测过程中以自动及时的方式表达,从而导致高产量的阳性克隆并节省大量时间。该方法已成功用于建立对人IgG具有高亲和力和特异性的单克隆scFv。
更新日期:2020-08-21
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