摘要
目的 研究一株自制的 c- erb B- 2单克隆抗体 A18在乳腺癌中表达的特性。方法 应用免疫组织化学 SP法、蛋白质印迹分析法和荧光激活的流式细胞分选检测技术检测 A 18在 6 35例乳腺癌组织、10 0例癌旁乳腺组织、不表达 c- erb B-2的 NIH /3T3(鼠成纤维细胞 )和 NE91(转表皮生长因子受体基因的 NR6鼠成纤维细胞 )细胞株及高表达 c- erb B- 2的 T6 -17(转 c- erb B- 2基因的 NIH /3T3细胞 )和 SKBR3(人乳腺癌细胞 )细胞株中的表达状况 ,并与市售进口 c- erb B- 2抗体 (MaximBiotech产品 )进行了平行对照研究。 A 18系采用细胞表面区域表位包埋法免疫小鼠制备而成。结果 A18阳性染色定位于细胞膜 ,部分伴微弱的细胞浆着色 ,无明显非特异性染色。 A18和进口抗体对 NIH/3T3、 NE91细胞均呈阴性 ,T6 - 17、 SKBR3细胞均呈阳性。在乳腺癌组织中 ,A18的阳性率为 6 0 .3%,明显高于癌旁乳腺组织的 5 .0 %。 A18与进口同类单抗的阴性、阳性及总符合率分别为 84.0 %、82 .8%及 83.2 %,与进口同类多抗的阴性、阳性及总符合率分别为 88.0 %、90 .2 %和 89.5 %。A18和进口同类单抗与乳腺癌临床病理特征的关系一致。经反复冻融 7次或 4℃保存 10个月 A18效价仍保持在 3μg/ml。结论 A18特异性强、定位准确、效价高?
? Objective To identify the immunohistochemical characteristics of a patent anti erbB2 antibody named A18, which was prepared by surface epitope masking technique in our laboratory. Method Four cell lines_NIH/3T3 (murine fibroblasts without c erbB 2 expression), T6 17 (NIH/3T3 fibroblasts transfected with c erbB 2 gene), NE91 (murine NR6 fibroblasts transfected with epithelial growth factor receptor and without c erbB 2 expression) and SKBR3 (a human breast cancer cell line overexpressing c erbB 2)_as well as 635 cases of human breast cancer tissue and 100 cases of adjacent breast tissue were detected immunohistochemically with A18, commercial monoclonal and multiclonal anti erbB2 antibodies (Maxim Biotech Inc). Western Blotting and flow cytometry analysis for A18 were also performed. Result A18 positive membrane staining was evident without non specific reaction. T6 17 and SKBR3 were positive for A18, while NIH/3T3 and NE91 were negative. A18 positive rate in breast cancer tissue was 60 3%, significantly higher than in the adjacent benign breast tissue (5 0%). The negative, positive and total coincidence rates of A18 with the commercial monoclonal antibody in breast cancer tissues were 84 0%, 82 8% and 83 2% respectively, and those of A18 with the commercial multiclonal antibody were 88 0%, 90 2% and 89 5% respectively. It also corresponded with the commercial monoclonal antibody in respect of clinicopathological features. A18 working concentration remained 3 μg/ml after 7 times of freeze thaw or storing at 4℃ for 10 months. Conclusion All these suggest its specificity, accuracy, high efficiency and stability in the detection of breast cancer.\;〔
出处
《中国组织化学与细胞化学杂志》
CAS
CSCD
2002年第1期14-19,115,共7页
Chinese Journal of Histochemistry and Cytochemistry
基金
国家科技攻关地方重大项目(No.99-D050)
