DU Chun-Hong, Wang Peng, Zhang Jian-Zhong, Tang Xue, Song Zhi-Zhong
Yunnan Institute for Endemic Diseases Control and Prevention, Dali 671000, China.
Xi Bao Yu Fen Zi Mian Yi Xue Za Zhi. 2011 Sep;27(9):985-8.
To prepare the monoclonal antibody of Pla with recombinant Pla (rPla) by hybridoma cell technology, which will lay the foundation for related research work.
Purified rPla was collected by washing repeatedly with urea, and BALB/c mice were immunized by them. Hybridoma cells were achieved by Sp2/0 cell fusion with mouse spleen cells from successfully immunized mice. Monoclonal antibody was screened by indirect ELISA and Western blots with rPla, natural crude Pla and GST respectively.
Three strains of hybridoma cells (named 15B8, 14H4 and 19A4 respectively) which secreted stably the monoclonal antibody of Pla were obtained. Their subclasses were IgG2a and IgG1 in heavy chains and κ chains in light chains. The ELISA titers of ascites were 10(6); respectively.Three of monoclonal antibody can react with natural crude Pla tested by western blots.
Monoclonal antibody of natural Pla of Yersinia pestis were successfully got, which has laid the foundation for further study of the Pla protein and development diagnosis reagent.
