Preparation and Identification of Monoclonal Antibody Specifically Against Human NT-proBNP

doi:10.13560/j.cnki.biotech.bull.1985.2016.06.021

Abstract

Abstract: This study aims to prepare and identify monoclonal antibodies specifically against human NT-proBNP. Balb/c mice was immunized with labeled recombinant protein NT-proBNP，then the spleen cells with high titer of the mice’s serum were fused with SP2/0 myeloma cells. Further，the positive hybridoma cell lines were established after HAT selection and subcloning by limiting dilution，ascites were prepared by injecting cells to mice’s abdomen，and monoclonal antibodies were purified by caprylic acid-ammonium sulfate precipitation method. Finally，basic functions of these monoclonal antibodies such as subclasses，titers，specificity，affinity constants were analyzed. Four positive hybridoma cell lines stably secreting human monoclonal antibodies of anti-NT-proBNP were screened out，and named as 1G12，1B2，3D5，and 2D11；1G12 and 1B2 belonged to IgG2a，2D11 to IgG1，and 3D5 to IgG2b. The titers of 1G12，1B2 and 3D5 reached 10⁶，while the titer of 2D11 only was 10⁴. The affinity constants of 1G12，1B2，3D5 and 2D11 were 9.32×10¹¹，1.07×10¹²，2.31×10¹²，and 6.33×10¹⁰，respectively. In conclusion，the monoclonal antibody of specifically against human NT-proBNP was prepared successfully.

Key words: NT-proBNP, monoclonal antibodies, Enzyme-linked immunosorbent assay

LIU Peng-zhan, HUANG Qi-ling, HE Xiao-wei, LI Wen-mei, ZHANG Wen-qi, ZHANG Sai. Preparation and Identification of Monoclonal Antibody Specifically Against Human NT-proBNP[J]. Biotechnology Bulletin, 2016, 32(6): 148-154.

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