Construction and identification of full human monoclonal antinuclear autoantibody
Jing Wu, Yang Li, Qiuyuan Han, Xin Yu, Wenping Kao, Wuqi Song, Aixia Zhai, Yujun Li, Aimei Li, Qingmeng Zhang, Chao Pan, Fengmin Zhang
Abstract
ObjectiveThis study is aimed to obtain autoantibodies from the peripheral blood lymphocytes of systemic lupus erythematosus (SLE) patients, and to screen the antinuclear antibody by indirect immunofluorescence identification.
MethodsThe technology of human B lymphocyte hybridoma was used in the present study for the preparation of the full human autoantibody derived from SLE patients.ELISA was performed to detect the concentration of the monoclonal antibodies in the positive hybridoma cell culture supernatant.Indirect immunofluorescence assay was performed to identify the antinuclear antibody by using HEp-2 cells as a basal substrate.
ResultsForty eight strains of full human monoclonal autoantibody secreting hybridoma cells strains were prepared, and the concentration in twelve strains is higher than 1 500 ng/mL with a ratio of 25%.Three types of antinuclear antibody were identified and screened from the full human monoclonal autoantibody library, and the immunofluorescence pattern was peripheral, speckled, and nuclear, respectively.
ConclusionThe preparation of full human monoclonal autoantibody and identification of antinuclear antibody could provide a experimental basis for the development of new monoclonal antibody drugs and for the study of researching the pathogenesis of SLE.
Key words:
Systemic lupus erythematosus; Autoantibody; Antinuclear antibody; Human B lymphocyte hybridoma technology
Contributor Information
Jing Wu
Department of Microbiology, Harbin Medical University, the Heilongjiang Key Laboratory of Immunity and Infection, Pathogenic Biology, Wu Lien-Teh Institute, Harbin 150081, China
Yang Li
Department of Radiotherapy, the Third Affiliated Hospital of Harbin Medical University, Harbin 150081, China
Qiuyuan Han
Department of Critical Care Medicine, the Second Affiliated Hospital of Harbin Medical University, Harbin 150081, China
Xin Yu
Department of Medical Laboratory Medicine, Affiliated Zhongshan Hospital of Dalian University, Dalian 116001, China
Wenping Kao
Department of Microbiology, Harbin Medical University, the Heilongjiang Key Laboratory of Immunity and Infection, Pathogenic Biology, Wu Lien-Teh Institute, Harbin 150081, China
Wuqi Song
Department of Microbiology, Harbin Medical University, the Heilongjiang Key Laboratory of Immunity and Infection, Pathogenic Biology, Wu Lien-Teh Institute, Harbin 150081, China
Aixia Zhai
Department of Microbiology, Harbin Medical University, the Heilongjiang Key Laboratory of Immunity and Infection, Pathogenic Biology, Wu Lien-Teh Institute, Harbin 150081, China
Yujun Li
Department of Microbiology, Harbin Medical University, the Heilongjiang Key Laboratory of Immunity and Infection, Pathogenic Biology, Wu Lien-Teh Institute, Harbin 150081, China
Aimei Li
Department of Microbiology, Harbin Medical University, the Heilongjiang Key Laboratory of Immunity and Infection, Pathogenic Biology, Wu Lien-Teh Institute, Harbin 150081, China
Qingmeng Zhang
Department of Microbiology, Harbin Medical University, the Heilongjiang Key Laboratory of Immunity and Infection, Pathogenic Biology, Wu Lien-Teh Institute, Harbin 150081, China
Chao Pan
Department of Microbiology, Harbin Medical University, the Heilongjiang Key Laboratory of Immunity and Infection, Pathogenic Biology, Wu Lien-Teh Institute, Harbin 150081, China
Fengmin Zhang
Department of Microbiology, Harbin Medical University, the Heilongjiang Key Laboratory of Immunity and Infection, Pathogenic Biology, Wu Lien-Teh Institute, Harbin 150081, China
