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目的制备获得人P53蛋白单克隆抗体,并应用于免疫组化检测。方法利用基因重组方法,将p53基因构建到原核质粒载体上并诱导表达,将收集获得的P53蛋白免疫小鼠,经细胞融合及亚克隆手段获得鼠源单克隆抗体,与商品化的抗体DO-1共同染色蜡块包埋的病理组织样本,比较二者的染色结果。结果在筛选获得的与DO-1检测结果相近的5株抗体内,取检测结果最接近的1株抗体5E10,经Protein A/G株纯化抗体,与收集获得的数例癌症组织蜡块进行免疫组化验证,发现结果均与DO-1的结果无明显差异。结论成功制备并筛选出1株人抗P53单克隆抗体,可用于免疫组化染色。
Objective To prepare monoclonal antibodies against human P53 protein and apply it to immunohistochemistry. METHODS: The p53 gene was constructed on the prokaryotic plasmid vector and induced by gene recombination method. The mouse P53 protein immunized mice were harvested to obtain murine monoclonal antibody by cell fusion and subcloning, 1 Co-staining of paraffin-embedded pathological tissue samples, comparing the two staining results. Results Five strains of antibodies similar to those detected by DO-1 were screened and one antibody closest to that of 5E10 antibody was obtained. The antibody was purified by Protein A / G strain and immunized with several pieces of cancer tissue wax collected Histochemistry and the results were found no significant difference with the DO-1 results. Conclusion One human anti-P53 monoclonal antibody was successfully prepared and screened for immunohistochemical staining.