Deng Yan, Lei Ming, Zhang Wei, Li Weiguo, Zhou Esheng, Liu Zhonglai, Qi Chao
Hubei Key Laboratory of Genetic Regulation and Integrative Biology, College of Life Science, Huazhong Normal University, Wuhan, People's Republic of China.
Hybridoma (Larchmt). 2011 Oct;30(5):491-3. doi: 10.1089/hyb.2011.0053.
Fpr3 is the third member of the FKBP (FK506 binding protein) family in yeast. In this study, the fpr3 gene from Saccharomyces cerevisiae was overexpressed and the protein product was purified using different methods. The recombinant Fpr3 fusion protein (rFpr3) was then used as antigen to immunize BALB/c mice for the production of monoclonal antibodies (MAb). Western blot and ELISA results indicated that rFpr3 had specific binding ability to the MAbs, and isotyping results classified the MAb as the subclass IgG1 by antibody. The MAbs obtained in this study will be used as a molecular chaperone to obtain Fpr3 crystals.
Fpr3是酵母中FKBP（FK506结合蛋白）家族的第三个成员。在本研究中，酿酒酵母的fpr3基因被过量表达，并且使用不同方法纯化了蛋白质产物。然后将重组Fpr3融合蛋白（rFpr3）用作抗原免疫BALB/c小鼠以产生单克隆抗体（MAb）。蛋白质免疫印迹和酶联免疫吸附测定结果表明，rFpr3对这些单克隆抗体具有特异性结合能力，并且通过抗体同种型鉴定结果将单克隆抗体归类为IgG1亚类。本研究中获得的单克隆抗体将用作分子伴侣以获得Fpr3晶体。
