Liu Tao, Liu Yan, Wang Ying, Hull Mitchell, Schultz Peter G, Wang Feng
Department of Chemistry and the Skaggs Institute for Chemical Biology, The Scripps Research Institute , La Jolla, California 92037, United States.
J Am Chem Soc. 2014 Jul 30;136(30):10557-60. doi: 10.1021/ja5042447. Epub 2014 Jul 18.
The bovine antibody (BLV1H12) which has an ultralong heavy chain complementarity determining region 3 (CDRH3) provides a novel scaffold for antibody engineering. By substituting the extended CDRH3 of BLV1H12 with modified CXCR4 binding peptides that adopt a β-hairpin conformation, we generated antibodies specifically targeting the ligand binding pocket of CXCR4 receptor. These engineered antibodies selectively bind to CXCR4 expressing cells with binding affinities in the low nanomolar range. In addition, they inhibit SDF-1-dependent signal transduction and cell migration in a transwell assay. Finally, we also demonstrate that a similar strategy can be applied to other CDRs and show that a CDRH2-peptide fusion binds CXCR4 with a K(d) of 0.9 nM. This work illustrates the versatility of scaffold-based antibody engineering and could greatly expand the antibody functional repertoire in the future.
具有超长重链互补决定区3（CDRH3）的牛抗体（BLV1H12）为抗体工程提供了一种新型支架。通过用采用β-发夹构象的修饰CXCR4结合肽替代BLV1H12的延伸CDRH3，我们生成了特异性靶向CXCR4受体配体结合口袋的抗体。这些工程化抗体以低纳摩尔范围内的结合亲和力选择性结合表达CXCR4的细胞。此外，它们在transwell测定中抑制SDF-1依赖性信号转导和细胞迁移。最后，我们还证明类似的策略可应用于其他互补决定区（CDR），并表明CDRH2-肽融合体以0.9 nM的解离常数（K(d)）结合CXCR4。这项工作说明了基于支架的抗体工程的多功能性，并可能在未来极大地扩展抗体功能库。
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