A humanized monoclonal antibody targeting secreted anterior gradient 2 effectively inhibits the xenograft tumor growth
A humanized monoclonal antibody targeting secreted anterior gradient 2 effectively inhibits the xenograft tumor growth
复制标题
靶向分泌前梯度2的人源化单克隆抗体有效抑制异种移植肿瘤生长
DOI：
10.1016/j.bbrc.2016.05.033
复制
发表时间：
2016-06-17
影响因子：
3.1
通讯作者：
Li, Dawei
中科院分区：
文献类型：
--
作者：
Guo, Hao;Chen, Hao;Li, Dawei
前梯度2（AGR 2）是潜在的抗肿瘤靶标，并且我们先前报道了与AGR 2特异性结合的鼠抗体18 A4。然而，人源化是在考虑临床使用之前克服免疫原性的必要条件，并且用于哺乳动物表达的优化载体对于随后的工业化生产也是必要的。在此，我们描述了阻断分泌的AGR 2活性的抗肿瘤人源化抗体。采用CDR移植技术和去免疫分析方法构建了18 A4的人源化抗体变体，并通过理化性质比较，筛选出18 A4 Hu I作为最佳的人源化候选抗体。小鼠移植瘤实验表明18 A4 Hu I能有效抑制移植瘤生长，AGR 2突变体抗体封闭表位分析表明18 A4 Hu I的抑制活性可能是通过阻断依赖于E60-H76和A86-E153氨基酸位点的AGR 2功能发挥的。此外，我们还设计了一个pHAb-FAST载体系统，用于人源化抗体哺乳动物表达载体的快速构建。在pHAb-FAST系统中，仅需两次重叠PCR反应即可快速构建18 A4 Hu I的表达载体。以AGR 2为靶点的18 A4 Hu I是一种很有前途的人源化抗肿瘤药物候选者，pHAb-FAST系统是一种有效的优化哺乳动物表达载体构建工具。我们的研究结果有望加速基于抗体的癌症治疗的发展。(C)2016 Elsevier Inc. All rights reserved.
Anterior Gradient 2 (AGR2) is a potential anti-tumor target and we previously reported a murine antibody 18A4 with specific binding to AGR2. However, humanization is a must to overcome immunogenicity before considering for clinical use and optimized vectors for mammalian expression are also necessary for following industrialized manufacture. Here, we describe an anti-tumor humanized antibody blocking secreted AGR2 activity. We employed the CDR grafting technique and deimmunization analysis to construct humanized antibody variants of 18A4, and 18A4Hu I was selected as the best humanization candidate, characterized by physical and chemical property comparison. Mouse xenograft study showed that 18A4Hu I could effectively inhibit the xenograft tumor growth, antibody blocking epitope analysis using AGR2 mutants indicated that the inhibition activity of 18A4Hu I is exerted probably through blocking the AGR2 functions which rely on the amino acid sites of E60-H76 and A86-E153. What's more, in this report, we also describe a pHAb-FAST vector system which is specifically designed for humanized antibody mammalian expression vector fast construction. With pHAb-FAST system, expression vector of 18A4Hu I could be quickly constructed only through twice overlapping PCR reactions. To our knowledge, AGR2-targeted 18A4Hu I is a promising humanized anti-tumor drug candidate, and pHAb-FAST system is a useful optimized mammalian expression vector construction tool. Our findings are supposed to accelerate the development of antibody-based cancer therapy. (C) 2016 Elsevier Inc. All rights reserved.