Affinity maturation of an TpoR targeting antibody in full-length IgG form for enhanced agonist activity.
Affinity maturation of an TpoR targeting antibody in full-length IgG form for enhanced agonist activity.
复制标题
全长 IgG 形式的 TpoR 靶向抗体的亲和力成熟可增强激动剂活性。
DOI：
10.1093/protein/gzy002
复制
发表时间：
2018
影响因子：
2.4
通讯作者：
Lerner Richard A
中科院分区：
文献类型：
--
作者：
Yang Zhuo;Du Mingjuan;Wang Wei;Xin Xiu;Ma Peixiang;Zhang Hongkai;Lerner Richard A
已经观察到，将scFv形式的抗体转化为全长IgG通常与亲和力的丧失有关。本文旨在通过建立一种应用酵母展示技术平台的整合亲和力成熟方法来解决这一问题。为了证明这一点，我们采用了一种名为3D 9的人血小板生成素受体靶向抗体，其先前从scFv-Fc融合蛋白形式的组合抗体文库中鉴定。我们已经观察到，当3D 9转化为全长IgG形式时，发生显著的效力损失。在这项研究中，我们测试了全长IgG的效力是否可以通过使用修饰的Fab酵母展示平台的3D 9的亲和力成熟来提高。针对具有预先设计的CDR多样性的Fab文库，设计了一种有效的CDR 3靶向突变策略。下一代测序也用于评价富集过程和研究抗体的序列-功能关系。鉴定出了亲和力提高、效价更高的变体。研究表明，我们在这里使用的策略是有效的优化全长IgG的亲和力和活性。
It has been observed that converting scFv formatted antibodies to full-length IgG often associates with loss of affinity. We aim to address this issue in this paper by establishing an integrated affinity maturation method applying yeast display technology platform. To demonstrate that, we employed a human thrombopoietin receptor targeting antibody named 3D9 which was identified previously from a combinational antibody library in scFv-Fc fusion protein form. We have observed that significant potency loss happened when 3D9 was transformed to full-length IgG form. In this study, we tested whether the potency of the full-length IgG can be improved by affinity maturation of 3D9 using a modified Fab yeast display platform. An efficient CDR3 targeted mutagenesis strategy was designed for Fab library with pre-designed CDR diversity. Next generation sequencing was also used for evaluation of the enrichment process and investigation of sequence-function relationship of the antibody. A variant with improved affinity and higher potency was identified. The study demonstrates that the strategy we used here are efficient for optimizing affinity and activity of full-length IgGs.