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目的 :制备抗PAGE 4mAb并鉴定其特异性及敏感性 ,应用该抗体在蛋白质水平上分析PAGE 4在正常与疾病组织中的表达 .方法 :以PAGE 4为抗原免疫BALB/c小鼠 ,利用杂交瘤技术建立稳定分泌抗PAGE 4mAb的杂交瘤细胞系 .采用WesternBlot及ELISA等方法鉴定抗体的特异性和敏感性 .应用正辛酸 饱和硫酸铵方法纯化抗体 .利用免疫组织化学方法对PAGE 4在正常与疾病组织中的表达进行分析 .结果 :获得 4株分泌特异性抗PAGE 4mAb的杂交瘤细胞株 ,mAb具有较高的特异性与敏感性 .在正常男女生殖系统组织如 :睾丸、前列腺、子宫中以及正常胰腺组织中有PAGE 4的表达 ,在前列腺癌、子宫内膜腺癌中PAGE 4高表达 .PAGE 4在蛋白质水平上的表达格局与以往有关PAGE 4在mRNA水平上的报道一致 .结论 :成功获得抗PAGE 4mAb ,为研究PAGE 4定位及功能提供了重要工具 .
Objective: To prepare anti-PAGE 4 mAb and identify its specificity and sensitivity, and to analyze the expression of PAGE 4 in normal and diseased tissues by using this antibody.Methods: BALB / c mice were immunized with PAGE 4 antigen, To establish a hybridoma cell line stably secreting anti-PAGE 4 mAb.Methods Western blot and ELISA were used to identify the specificity and sensitivity of the antibody.Antibodies were purified by saturated ammonium sulfate-n-octanoate.Proteinase- The results showed that 4 McAbs secreting specific anti-PAGE 4 mAb were obtained, and mAb was highly specific and sensitive.In the normal male and female reproductive system tissues such as testis, prostate and uterus PAGE4 expression in normal pancreatic tissue and high expression in prostate cancer and endometrial adenocarcinoma PAGE 4. The expression pattern of PAGE 4 at the protein level is consistent with previous reports on the mRNA level of PAGE 4. Conclusion: The successful anti-PAGE 4 mAb provided an important tool for studying PAGE 4 localization and function.