A Streamlined Approach for the Construction of Large Yeast Surface Display Fab Antibody Libraries
A Streamlined Approach for the Construction of Large Yeast Surface Display Fab Antibody Libraries
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DOI：
10.1007/978-1-4939-8648-4_8
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发表时间：
2018-01-01
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影响因子：
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通讯作者：
Zielonka, Stefan
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文献类型：
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作者：
Krah, Simon;Grzeschik, Julius;Zielonka, Stefan
酵母表面展示是一种多功能的抗体发现平台技术。然而，抗体Fab文库的构建通常是繁琐的多步骤过程，其涉及在不同的单倍体酵母菌株中产生重链以及轻链展示质粒，然后进行酵母交配。在这里，我们提出了一个集中的一步金门克隆的酵母表面展示Fab库，允许同时引入重链和轻链可变区到一个单一的展示载体的生成方法。因此，可以显著减少文库生成所需的总时间以及材料。
Yeast surface display is a versatile platform technology for antibody discovery. Nevertheless, the construction of antibody Fab libraries typically is a tedious multistep process that involves the generation of heavy chain as well as light chain display plasmids in different haploid yeast strains followed by yeast mating. Here, we present a focused one-step Golden Gate cloning approach for the generation of yeast surface display Fab libraries that allows for simultaneous introduction of heavy-chain and light-chain variable regions into one single display vector. Thereby, the overall time as well as the materials needed for library generation can be reduced significantly.