鼠抗人DCP单克隆抗体的类型转换与表达  

Class switching and expression of a mouse anti-human DCP antibody

作　　者：廖凯 张智萍 丁笠 张新跃 LIAO Kai;ZHANG Zhiping;DING Li;ZHANG Xinyue(College of Bioscience and Biotechnology,Yangzhou University,Yangzhou 225009,China)

机构地区：[1]扬州大学生物科学与技术学院,江苏扬州225009

出　　处：《扬州大学学报（农业与生命科学版）》2024年第3期72-81,共10页Journal of Yangzhou University：Agricultural and Life Science Edition

基　　金：江苏省研究生科研创新计划项目(KYCX21-3206)。

摘　　要：为建立抗体类型转换和表达体系,以最新制备的1株IgM类抗体——鼠抗人DCP单克隆抗体7C2为研究对象,首先从7C2杂交瘤细胞中克隆IgM轻重链可变区,再将IgM可变区序列与鼠IgG1恒定区序列融合克隆至真核表达载体,并转入293细胞系中。利用蛋白质免疫印迹技术,成功检测到IgG1抗体在细胞内的表达并分泌至培养上清。在进一步优化密码子、宿主细胞和信号肽等影响抗体表达的因素后,建立了双质粒瞬转和基于双顺反子表达方式的稳转表达体系,可用于将鼠源IgM类抗体转换为IgG类抗体并进行表达。该研究为拓展非IgG类抗体的应用范围提供重要途径,同时为提高重组抗体在真核体系中的表达效率提供参考。In this study,we used a recently-prepared mouse anti-human DCP monoclonal antibody,7C2,of IgM class as a research object to establish a system for class switching and expression of antibody.The variable regions of light and heavy chains of 7C2antibody were cloned from hybridoma cells.Then,we fused the variable region sequences of 7C2with the constant region sequences of mouse IgG1antibody into a eukaryotic expression vector and transferred the recombinant plasmid into 293cell line.Western blot analysis showed that recombinant IgG1antibody was successfully expressed in the cells and secreted into culture supernatant.After further optimizing the factors affecting the expression efficiency of antibodies,such as codons,host cells and signal peptides,we established a double-plasmid transient expression system and a bicistronic expression system,which can be used to convert murine IgM into IgG antibodies and express them.This study provides an important way to expand the application of non-IgG type antibodies and also a reference for improving the expression efficiency of recombinant antibodies in eukaryotic system.

关 键 词：单克隆抗体 基因工程 抗体类型转换 表达条件优化 

分 类 号：S852.4[农业科学—基础兽医学] Q789[农业科学—兽医学]