生产新型抗缪勒氏管激素受体 2 多肽靶向抗体并诱导卵巢癌细胞产生细胞毒性。

IF 4.1 4区 医学 Q2 IMMUNOLOGY
Scandinavian Journal of Immunology Pub Date : 2025-01-01 Epub Date: 2024-11-25 DOI:10.1111/sji.13426
Çağrı Şakalar, Büşra Kurt, Sedat Sezen, Savaş Kaya
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引用次数: 0

摘要

卵巢癌通常在晚期才被诊断出来。针对卵巢癌抗原的单克隆抗体(MAbs)已用于临床。抗缪勒氏管激素受体2型(AMHR2)是一种在卵巢癌中高度表达的受体,也是一种潜在的免疫疗法靶抗原。研究人员对 AMHR2 的胞外结构域进行了三维结构和理化性质分析,并确定了 3 个肽序列(肽 1、肽 7 和肽 11)作为靶标。进行了 MAb 生产规程,获得了 6 个对肽具有高亲和力的 MAb 克隆。P3B1、P10A10、P10B6 和 P2A6 克隆针对肽 11(P11),P2C9 针对 P7,P6C5 针对 P1。P2A6 的抗体同种型为 IgG2a,其他抗体同种型为 IgG1。酶联免疫吸附试验(ELISA)分析了 MAb 与原生重组蛋白(AMHR2-Fc)的结合情况,Western 印迹和免疫荧光染色分析了 MAb 与 SKOV-3 卵巢癌细胞表达的 AMHR2 的结合情况。P3B1 与原生蛋白(AMHR2-Fc)的亲和力较强,P10A10、P10B6 和 P2C9 与原生蛋白(AMHR2-Fc)的亲和力中等。P3B1 和 P2C9 在 Western 印迹分析中显示出很强的结合力。克隆在免疫荧光染色中显示出中等程度的结合力。使用 MAbs 和转染的 SKOV-3 细胞进行了补体依赖性细胞毒性(CDC)实验。P3B1 诱导了明显的 CDC。对 P3B1 MAb 的可变区进行了测序。总之,针对 AMHR2 的三个不同区域制备了 MAb。其中一个克隆可诱导卵巢癌细胞产生细胞毒性,其序列已被确定,将来可用作人源化治疗 MAb。
本文章由计算机程序翻译,如有差异,请以英文原文为准。
Production of novel peptide-targeting antibodies for anti-Müllerian hormone receptor 2 and induction of cytotoxicity in ovarian cancer cells.

Ovarian cancer is generally diagnosed at late stages. Monoclonal antibodies (MAbs) targeting antigens in ovarian cancer are used in the clinic. Anti-Müllerian hormone receptor type 2 (AMHR2) is a receptor highly expressed in ovarian cancer and it is a potential target antigen for immunotherapy. Extracellular domain of AMHR2 was analysed in terms of 3D structure and physicochemical properties, and 3 peptide sequences (Peptides 1, 7 and 11) were determined as targets. MAb production protocol was performed, and 6 MAb clones showing high affinity for peptides were obtained. P3B1, P10A10, P10B6 and P2A6 clones were for peptide 11 (P11), P2C9 was for P7, and P6C5 was for P1. Antibody isotype of P2A6 was IgG2a and the others were of IgG1 isotype. MAb binding to the native recombinant protein (AMHR2-Fc) was analysed by enzyme-linked immunosorbent assay (ELISA) and MAb binding to AMHR2 expressed by SKOV-3 ovarian cancer cells was analysed by western blot and immunofluorescent staining. P3B1 showed strong, P10A10, P10B6 and P2C9 showed medium affinity for the native protein (AMHR2-Fc). P3B1 and P2C9 showed strong binding in western blot analysis. Clones showed moderate binding in immunoflorescent staining. A complement dependent cytotoxicity (CDC) experiment was conducted using MAbs and transfected SKOV-3 cells. P3B1 induced a significant CDC. Variable regions of P3B1 MAb were sequenced. In conclusion, MAbs for three different regions of AMHR2 were produced. One clone was shown to induce cytotoxicity in ovarian cancer cells and its sequence was determined for future use as a humanised therapeutic MAb.

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来源期刊
CiteScore
7.70
自引率
5.40%
发文量
109
审稿时长
1 months
期刊介绍: This peer-reviewed international journal publishes original articles and reviews on all aspects of basic, translational and clinical immunology. The journal aims to provide high quality service to authors, and high quality articles for readers. The journal accepts for publication material from investigators all over the world, which makes a significant contribution to basic, translational and clinical immunology.
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