MUC1核心肽多克隆抗体的制备及鉴定

Preparation and Identification of Polyclonal Antibody Against MUC1 Core Peptide

目的 制备MUC1核心肽多克隆抗体 ,用于靶向MUC1肿瘤治疗的研究。方法 以 8R -MUC1核心肽 6重复序列重组蛋白 (8R MUCPT)为抗原免疫家兔 ,以ELISA法确定抗MUC1多克隆抗体效价 ,以Westernblot和ELISA阻断试验鉴定抗体特异性。结果 MUC1核心肽多克隆抗体效价为 1∶2 5 6 0 0 0 ,能与MUC1核心肽特异性结合。结论 利用原核表达的 8R -MUCPT蛋白 ,可有效地诱导免疫动物产生MUC1核心肽特异性抗体 ,为MUC1核心肽靶向肿瘤特异性免疫研究提供了有力的实验材料。

Objective To prepare anti-MUC1 core peptide polyclonal antibody used as target in immunotherapy of tumors.Methods Immunize rabbits with recombinant 8R-MUC1 core peptide protein with s ix tandem repeat sequence,and determine the titer of polyclonal antibody against MUC1 by ELISA.The specificity of antibody were identified by Western blot and b locking ELISA separately.Results The polyclonal antibody agains t MUC1,with a titer of 1∶256 000,bound specifically to MUC1 core peptide. Conclusion The 8R-MUCPT protein expressed in prokaryotic system effe ctively induced specific antibody against MUC1 core peptide in immunized animals .It provided an important experimental materials for the study on tumor-targeti ng MUC1 core peptide in immunotherapy. [