Abstract:Abstract: Parathyroid hormone (PTH) is a straight-chain polypeptide hormone containing only 84 amino acids (aa), and is one of the main hormones regulating calcium and phosphorus metabolism. The hormone is used to evaluate the prognosis of patients with bone conversion, kidney disease and hemodialysis. In order to prepare monoclonal antibodies against PTH and establish a rapid detection method of PTH, the artificially synthesized PTH peptide was coupled to keyhole limpet hemocyanin (KLH), and the coupled PTH was used to immunize BALB/c mice. After cell fusion and hybridoma cell screening, a total of 5 monoclonal antibodies that stably secreted anti-PTH were obtained, and monoclonal antibodies were prepared and purified. Then, the antibody pairing screening was performed by phalanx checkerboard titration. The 9B6 and 22E8 antibodies were used as the capture antibody and HRP-labeled antibody, respectively, and the double-antibody sandwich enzyme linked immunosorbent assay (dsELISA) method for detecting PTH was established. After optimization, the detection sensitivity for dsELISA method reached 0.14 ng/mL, and the method also has a good linearity within the range of 0.5~8.0 ng/mL, with its R2 about 0.985 8 and precision below 10%. In conclusion, monoclonal antibodies against PTH were successfully prepared, and an ELISA detection method for PTH was established, which had good linear correlation, accuracy and precision, and could be used for the detection of PTH content.
引用本文:
王 辉, 徐 叶, 汪子力, 彭惠芳, 曾 璇, 李明勇, 刘利辉, 王庆林, 刘如石. PTH单克隆抗体的制备与初步应用[J]. 生命科学研究, 2022, 26(6): 509-514.
WANG Hui, XU Ye, WANG Zi-li, PENG Hui-fang, ZENG Xuan, LI Ming-yong, LIU Li-hui, WANG Qing-lin, LIU Ru-shi. Preparation and Preliminary Application of Monoclonal Antibodies Against PTH. Life Science Research, 2022, 26(6): 509-514.