Single-chain variable fragment (scFv) is the most common format for phage display antibody library. The isolated scFvs need to be reformatted to full-length IgGs for further characterization. High throughput reformatting of scFv to IgG without disrupting VH–VL pairing is of great demanding for exhaustive screening of all antibodies in IgG format. Herein, we developed a strategy based on the overlap extension PCR in emulsion to reformat scFv to IgG while maintain the accuracy and complexity of variable region pairing. Using CD40 as an example target, we reformatted phage display derived CD40 binding scFv library to IgG mammalian display library and isolated high affinity CD40 binding IgGs. This robust and reliable antibody reformatting approach could be integrated into any phage display based antibody drug discovery.

中文翻译：

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通过一步乳液PCR将噬菌体展示的抗体片段对IgG的高通量重格式化

单链可变片段（scFv）是噬菌体展示抗体库的最常见格式。分离的scFv需要重新格式化为全长IgG，以进行进一步表征。在不破坏VH–VL配对的情况下，将scFv重整为IgG的高通量对彻底筛查IgG格式的所有抗体都提出了很高的要求。本文中，我们开发了一种基于乳剂中重叠延伸PCR的策略，可将scFv重新格式化为IgG，同时保持可变区配对的准确性和复杂性。使用CD40作为示例靶标，我们将噬菌体展示衍生的CD40结合scFv文库重新格式化为IgG哺乳动物展示文库，并分离了高亲和力的CD40结合IgG。这种强大而可靠的抗体重新格式化方法可以集成到任何基于噬菌体展示的抗体药物发现中。