Rational design of peptides for identification of linear epitopes and generation of neutralizing monoclonal antibodies against DKK2 for cancer therapy.
Rational design of peptides for identification of linear epitopes and generation of neutralizing monoclonal antibodies against DKK2 for cancer therapy.
复制标题
合理设计肽，用于鉴定线性表位并生成针对 DKK2 的中和单克隆抗体，用于癌症治疗。
DOI：
10.1093/abt/tbaa004
复制
发表时间：
2020
影响因子：
--
通讯作者：
Sun,Le
中科院分区：
文献类型：
--
作者：
Zhao,Rongqing;Xiao,Qian;Li,Maohua;Ren,Wenlin;Xia,Chenxi;Liu,Xudong;Li,Yingzi;Tan,Tan;Wu,Dianqing;Sun,Le
Dickkopf相关蛋白2（DKK 2）是Wnt信号通路中Dickkopf家族的成员。最近，我们发现针对DKK 2的抗体可以激活肿瘤中的自然杀伤（NK）和CD8+ T细胞，并抑制肿瘤生长。在本文中，我们报告的合理设计的肽鉴定线性表位和中和抗DKK 2单克隆抗体的产生。为了打破免疫耐受，我们设计并化学合成了六个对应于DKK 2不同区域的多肽作为免疫原，发现其中五个多肽可以产生与活性重组人DKK 2蛋白结合的小鼠多克隆抗体。用两种不同的多肽（34KLNSIKSSL42和240KVWKDATYS248）与匙孔血蓝蛋白（KLH）偶联免疫小鼠，成功地制备了抗人DKK 2的中和性单克隆抗体（5F8和1A10）。单克隆抗体不仅在体外消除DKK 2对Wnt信号传导的抑制，而且在体内抑制肿瘤生长。目前，这两种mAb正在进行人源化，作为免疫治疗候选物，并可能为治疗人类癌症提供新的药物。
Dickkopf-related protein 2 (DKK2)is a member of the Dickkopf family in Wnt signaling pathway. Recently, we found that antibodies against DKK2 could activate natural killer (NK) and CD8+ T cells in tumors and inhibit tumor growth. In this paper, we report the rational design of peptides for identification of linear epitopes and generation of neutralizing monoclonal anti-DKK2 antibodies. To break the immune tolerance, we designed and chemically synthesized six peptides corresponding to different regions of DKK2 as immunogens and found five of them could generate mouse polyclonal antibodies that can bind to the active recombinant human DKK2 protein. Neutralizing mouse monoclonal antibodies (5F8 and 1A10) against human DKK2 were successfully developed by immunizing the mice with two different peptides (34KLNSIKSSL42and240KVWKDATYS248) conjugated to Keyhole limpet hemocyanin (KLH). The monoclonal antibodies not only abolish DKK2’s suppression of Wnt signaling in vitro but also inhibits tumor growth in vivo. Currently, those two mAbs are undergoing humanization as immunotherapy candidates and may offer a new drug for treatment of human cancers.