title: Antigenic assessment of a recombinant human CD90 protein expressed in prokaryotic expression system.
abstract: 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.
authors: Department of Microbiology, Faculty of Advanced Sciences & Technology, Pharmaceutical Sciences Branch, Islamic Azad University, Tehran, Iran.
authors: Narges
authors: N
authors: Yousefi-Rad
authors: National Cell Bank of Iran, Pasteur Institute of Iran, Tehran, Iran.
authors: Mohammad Ali
authors: MA
authors: Shokrgozar
authors: Biotechnology Research Center, Pasteur Institute of Iran, Tehran, Iran.
authors: Mahdi
authors: M
authors: Behdani
authors: National Cell Bank of Iran, Pasteur Institute of Iran, Tehran, Iran.
authors: Shima
authors: S
authors: Moradi-Kalbolandi
authors: Department of Hepatitis and AIDs, Pasteur Institute of Iran, Tehran, Iran.
authors: Mahdieh
authors: M
authors: Motamedi-Rad
authors: National Cell Bank of Iran, Pasteur Institute of Iran, Tehran, Iran. Electronic address: habibi_m@pasteur.ac.ir.
authors: Mahdi
authors: M
authors: Habibi-Anbouhi
mesh_terms: D000818:Animals; D000906:Antibodies / Q000276:immunology; D000937:Antigen-Antibody Reactions; D001483:Base Sequence; D003001:Cloning, Molecular; D004797:Enzyme-Linked Immunosorbent Assay; D004926:Escherichia coli / Q000235:genetics*; D005260:Female; D006801:Humans; D008969:Molecular Sequence Data; D017434:Protein Structure, Tertiary; D011817:Rabbits; D011994:Recombinant Proteins / Q000737:chemistry / Q000235:genetics / Q000276:immunology; D018800:Thy-1 Antigens / Q000737:chemistry / Q000235:genetics* / Q000276:immunology*
chemical_list: D000906:Antibodies; D011994:Recombinant Proteins; D018800:Thy-1 Antigens
keywords: Escherichia coli (E. coli); Human CD90 (Thy-1); Prokaryotic expression system; Recombinant protein