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首页> 外文期刊>Applied Microbiology and Biotechnology >Construction of a large phage display antibody library by in vitro package and in vivo recombination
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Construction of a large phage display antibody library by in vitro package and in vivo recombination

机译：通过体外包装和体内重组构建大型噬菌体展示抗体库

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摘要

Capacity and diversity are extremely important to the quality of various phage display libraries. In this work, λ phage-based in vitro package was applied to construct a filamentous phage display antibody library so as to enlarge its capacity and introduce more sequence diversity in the final library. In vivo recombination via Cre recombinase/lox sites was also exploited to create VH/VL combination diversity based on multivalent package of λ phage packaging extracts on phagemid DNA concatemers. The library constructed with 10 μg concatenated phagemid DNA and ten vials of λ phage packaging extracts was calculated to contain 1.40×1010 independent clones. Higher capacity can be easily achieved when more materials are consumed. This strategy is somewhat more efficient than prior methods.
机译：容量和多样性对于各种噬菌体展示库的质量极为重要。在这项工作中，基于λ噬菌体的体外包装被用于构建丝状噬菌体展示抗体文库，以扩大其容量并在最终文库中引入更多的序列多样性。还通过基于噬菌粒DNA连接体的λ噬菌体包装提取物的多价包装，利用Cre重组酶/ lox位点进行体内重组以产生VH / VL 组合多样性。计算了由10μg串联噬菌体DNA和十个小瓶的λ噬菌体包装提取物构建的文库，其中包含1.40×1010 独立克隆。当消耗更多材料时，可以轻松实现更高的容量。该策略比现有方法更有效。

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