Platform for high-throughput antibody selection using synthetically-designed antibody libraries.
Platform for high-throughput antibody selection using synthetically-designed antibody libraries.
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DOI：
10.1016/j.nbt.2015.11.005
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发表时间：
2016-09-25
影响因子：
5.4
通讯作者：
Kiss MM
中科院分区：
文献类型：
--
作者：
Batonick M;Holland EG;Busygina V;Alderman D;Kay BK;Weiner MP;Kiss MM
合成的人源化抗体库通常是通过随机结合抗体高变区中多个位置的变化而产生的。虽然这些文库具有很大的理论多样性(>；1020)，但通过转化大肠杆菌可以实现的实用多样性限制在1010左右。为了限制实际的多样性，使其序列更接近于自然人类抗体的多样性，我们使用完全预定义的互补决定区(CDR)生成了单链抗体噬菌体文库。我们已经使用这个文库来筛选针对四个人类蛋白靶标的新抗体，并证明了在早期选择轮中识别六个CDR中每一个的丰富序列可以用来重建对靶标具有选择性的共识抗体。
Synthetic humanized antibody libraries are frequently generated by random incorporation of changes at multiple positions in the antibody hypervariable regions. Although these libraries have very large theoretical diversities (>1020), the practical diversity that can be achieved by transformation of E. coli is limited to about 1010. To constrain the practical diversity to sequences that more closely mimic the diversity of natural human antibodies, we generated a scFv phage library using entirely pre-defined complementarity determining regions (CDR). We have used this library to select for novel antibodies against four human protein targets and demonstrate that identification of enriched sequences at each of the six CDRs in early selection rounds can be used to reconstruct a consensus antibody with selectivity for the target.