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粒细胞集落刺激因子(G-CSF)是一种造血调控因子,为鉴定、检测和纯化人G-CSF,制备了四株特异识别人G—CSF的单克隆抗体(单抗,McAb)。1 材料和方法1.1 rhG-CSF和rhG-CSF融合蛋白 均由本实验室克隆、表达和纯化。纯化后的样品经SDS—PAGE鉴定,纯度大于95％。1.2 单抗制备 以rhG-CSF融合蛋白为抗原,按常规方法制备单抗。1.3 单抗识别抗原表位鉴定试验 用15μg/ml的抗原包被检测板(50μl/孔),对杂交瘤细胞的培养上清及其系列倍比稀释液进行间接ELISA检测,找出使抗原饱含的各McAb的最大稀释度。
G-CSF is a hematopoietic regulatory factor. To identify, detect and purify human G-CSF, four monoclonal antibodies (mAb, McAb) that specifically recognize human G-CSF were prepared. 1 Materials and methods 1.1 rhG-CSF and rhG-CSF fusion protein were cloned from our laboratory, expression and purification. Purified samples were identified by SDS-PAGE with a purity greater than 95%. 1.2 monoclonal antibody rhG-CSF fusion protein as antigen, according to the conventional method of preparation of monoclonal antibody. 1.3 Identification of monoclonal antibody epitope recognition test With 15μg / ml antigen coated detection plate (50μl / well), the hybridoma cell culture supernatant and its serial dilutions of indirect ELISA test to find out the full antigen The maximum dilution of each McAb.