关键词: 人BPI23, 杂交瘤细胞, 单克隆抗体

Abstract: Objective　Preparation of anti-human BPI23 monoclonal antibody with hybridoma technique and preliminary analysis of the application. Methods The immunized mouse spleen cells and mouse myeloma cells were fused by routine method;the methods of indirect ELISA and Western-blot were used to screen the hybridoma cells which secreted monoclonal antibody;the positive clones were subcloned for three times with limited dilution method to acquire hybridoma cells which secreted anti-human BPI23 monoclonal antibody stably and efficiently;the amplified hybridoma cells were injected into abdominal cavity of mouse and the ascites was collected;the antibodies in ascites were purified,the type,class and subclass of antibodies were detected;the specificity of the antibody was analyzed by Western-blot;the titer of the monoclonal antibody was detected by indirect ELISA;the normal human neutrophil and mononuclear cells were separated and purified and were used for smear making,anti-human BPI23 monoclonal antibody was used to detect by immunohistochemistry. Results　Three hybridoma cell lines(1B4、9C12 and 2H11)secreting anti-human BPI23 monoclonal antibody stably and efficiently were acquired,The anti-human BPI23 monoclonal antibodies secreting by hybridoma cells belonged to kappa opioid IgM class,kappa opioid IgG1 subclass and kappa opioid IgG1 subclass respectively;the titer was 1.28×105、1.28×105 and 4.1×106 respectively,the content of purified antibodies was 0.208g/L、2.03g/L and 3.88g/L respectively;Western-blot showed that the monoclonal antibodies in ascites of tumor-bearing mice can recognize human BPI23 protein prepared in our lab and human BPI55 standards saled in the market specifically as well,but can't bind mouse BPI25 and human LBP;in immunohistochemistry, the 1B4、9C12 and 2H11 monoclonal antibodies can recognize BPI of human neutrophil specifically. Conclusion　The monoclonal antibodies against human BPI23 were prepared successfully , which lay a foundation for later establishment of BPI detecting kit.

Key words: human BPI23, hybridoma cell, monoclonal antibody