Class switching and expression of a mouse anti-human DCP antibody
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 7C2 antibody were cloned from hybridoma cells.Then,we fused the variable region sequences of 7C2 with the constant region sequences of mouse IgG1 antibody into a eukaryotic expression vector and transferred the recombinant plasmid into 293 cell line.Western blot analysis showed that recombinant IgG1 antibody was successfully expressed in the cells and secreted into culture supernatant.After further optimizing the factors affecting the expression efficiency of anti-bodies,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 ex-pression efficiency of recombinant antibodies in eukaryotic system.
monoclonal antibodygenetic engineeringantibody class switchoptimization of expression conditions
国家科技期刊平台
NSTL
万方数据
