Antigenic assessment of a recombinant human CD90 protein expressed in prokaryotic expression system

•We expressed the extra-membrane domain of human CD90 (exCD90) antigen in E. coli.•The epitope integrity of purified recombinant antigen was confirmed.•An ELISA was performed using 5E10 anti-CD90 mAb for binding study.•We produced a rabbit anti-exCD90 polyclonal antibody which stained CD90+ cells.•The exCD90 had an appropriate antigenic integrity for antibody production. Cluster of Differentiation 90 (CD90, Thy-1) has been proposed as one of the most important biomarkers in several cancer cells including cancer stem cells (CSCs). CD90 is considered as a potential normal stem cell and CSCs biomarker and also has been identified in lung cancer stem cells, hepatocellular carcinoma cells and high-grade gliomas. Using eukaryotic host systems involves complex procedures and frequently results in low protein yields. The expression of recombinant proteins in Escherichia coli is comparatively easier than eukaryotic host cells. The potential of large scale production of recombinant protein has made this system an economic production platform. In this study we expressed the extra-membrane domain of human CD90 (exCD90) antigen (Gln15–Cys130) in E. coli expression host cells. The epitope integrity of purified recombinant antigen was confirmed by antibody–antigen interaction using 5E10 anti-CD90 monoclonal antibody and binding study through ELISA and florescent staining of CD90+ cells in a flow cytometry experiment.