title: Exploitation of human CD99 expressing mouse myeloma cells as immunogen for production of mouse specific polyclonal antibodies.
abstract: In this study, we describe the application of a molecular biology technique for the production of mouse polyclonal antibodies (pAbs) specific to human cell surface molecules. Production of the pAb specific to the human CD99 surface molecule was used as the study model. The retroviral expression system was employed to generate human CD99 expressing mouse myeloma cells. After cell sorting and single cell cloning, a myeloma clone which stably expressed high levels of human CD99 on its surface was established. The human CD99 expressing mouse myeloma cells were then used as the immunogen for immunization of BALB/c mice. As endogenous proteins of mouse myeloma cells possess self-non-immunogenicity for BALB/c mice, after immunization, only the expressed human CD99 molecules induce antibody response. After three immunizations, high titers of mouse anti-CD99 pAbs were successfully produced. The produced pAb specifically reacted to both recombinant human CD99 and native CD99 molecules expressed on human blood cells. The established technology is simple and valuable for the production of pAbs specific to human CD99 membrane proteins which can be used for characterization of the CD99 molecule.
authors.affiliation: Division of Clinical Immunology, Department of Medical Technology, Faculty of Associated Medical Sciences, Chiang Mai University, Chiang Mai 50200, Thailand.
authors.forename: Nuchjira
authors.initials: N
authors.lastname: Takheaw
authors.affiliation: Division of Clinical Immunology, Department of Medical Technology, Faculty of Associated Medical Sciences, Chiang Mai University, Chiang Mai 50200, Thailand; Biomedical Technology Research Center, National Center for Genetic Engineering and Biotechnology, National Science and Technology Development Agency at the Faculty of Associated Medical Sciences, Chiang Mai University, Chiang Mai 50200, Thailand.
authors.forename: Witida
authors.initials: W
authors.lastname: Laopajon
authors.affiliation: Division of Clinical Immunology, Department of Medical Technology, Faculty of Associated Medical Sciences, Chiang Mai University, Chiang Mai 50200, Thailand.
authors.forename: Kantinan
authors.initials: K
authors.lastname: Chuensirikulchai
authors.affiliation: Division of Clinical Immunology, Department of Medical Technology, Faculty of Associated Medical Sciences, Chiang Mai University, Chiang Mai 50200, Thailand; Biomedical Technology Research Center, National Center for Genetic Engineering and Biotechnology, National Science and Technology Development Agency at the Faculty of Associated Medical Sciences, Chiang Mai University, Chiang Mai 50200, Thailand.
authors.forename: Watchara
authors.initials: W
authors.lastname: Kasinrerk
authors.affiliation: Division of Clinical Immunology, Department of Medical Technology, Faculty of Associated Medical Sciences, Chiang Mai University, Chiang Mai 50200, Thailand; Biomedical Technology Research Center, National Center for Genetic Engineering and Biotechnology, National Science and Technology Development Agency at the Faculty of Associated Medical Sciences, Chiang Mai University, Chiang Mai 50200, Thailand. Electronic address: supansa_pata@hotmail.com.
authors.forename: Supansa
authors.initials: S
authors.lastname: Pata
mesh_terms: D000071443:12E7 Antigen / Q000096:biosynthesis / Q000235:genetics / Q000276:immunology; D000818:Animals; D058846:Antibodies, Monoclonal, Murine-Derived / Q000276:immunology*; D006801:Humans; D007114:Immunization; D051379:Mice; D008807:Mice, Inbred BALB C; D009101:Multiple Myeloma / Q000235:genetics / Q000276:immunology / Q000378:metabolism
chemical_list: D000071443:12E7 Antigen; D058846:Antibodies, Monoclonal, Murine-Derived; C061828:CD99 protein, human
keywords: Cell surface molecule; Mammalian expression; Mouse immunization; Polyclonal antibody; Recombinant protein