全人单克隆抗核抗体的制备和鉴定

Construction and identification of full human monoclonal antinuclear autoantibody

目的本研究旨在从系统性红斑狼疮（systemmiclupuserythematosus，SLE）患者外周血淋巴细胞中获得自身抗体，并从中鉴定筛选出抗核抗体。方法利用人B细胞杂交瘤技术制备SLE患者来源的全人单克隆自身抗体，采用酶联免疫吸附实验（enzyme-linkedimmunosorbenkassay，ELISA）方法测定阳性杂交瘤细胞上清抗体浓度，并以HEp-2细胞为底物，采用间接免疫荧光法鉴定筛选出抗核抗体。结果共获得48株稳定分泌IgG型全人单克隆自身抗体的杂交瘤细胞株，其中12种细胞上清抗体浓度大于1500ng／mL，占25％，通过免疫荧光鉴定从中筛选出3种抗核抗体，荧光核型分别为周边型、斑点型和核仁型。结论全人单克隆自身抗体的制备及抗核抗体的筛选鉴定，为开发新型单克隆抗体药物以及研究SLE的致病机制奠定了良好的实验基础。

Objective This study is aimed to obtain autoantibodies from the peripheral blood lympho- cytes of systemic lupus erythematosus （SLE） patients, and to screen the antinuclear antibody by indirect immu- nofluorescence identification. Methods The 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 per- formed to detect the concentration of the monoclonal antibodies in the positive hybridoma cell culture superna- tant. Indirect assay was performed to identify the antint^clear antibody by using HEp-2 cells as a basal substrate. Results Forty 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 autoanti-body library, and the immunofluorescence pattern was peripheral, speckled, and nuclear, respectively. Conclu- sion The 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 researc- hing the pathogenesis of SLE.