自制抗人雌激素受体单克隆抗体的鉴定及临床应用研究

Appraisal and Clinical Application of Self made Monoclonal Antibody against Estrogen Receptor

目的：对自制抗人雌激素受体Ｄ段合成肽（ｈＥＲＤ）的单克隆抗体（ＭｃＡｂ）进行Ｗｅｓｔｅｒｎ免疫印迹特异性分析，显示自制单抗与野生型ＥＲ有特异性反应。方法：对５１例乳腺癌组织石蜡切片或冰冻切片进行免疫组化（ＩＨＣ）检测，并与葡聚糖包裹活性炭吸附法（ＤＣＣ）进行比较，符合率９２％（４７／５１）。间接荧光免疫反应检测自制抗ＥＲ单抗对ＭＣＦ－７培养细胞的荧光标记为阳性，以已知ＥＲ检测值的三个不同石蜡组织为抗原，鉴定自制单抗可否用于临床流式细胞术（ＦＣＭ）检测。结果：其ＥＲ的阳性标记率及荧光道数均与已知ＩＨＣ检测结果相符。结论：提示自制单抗也可用于流式免疫学方法检测ＥＲ。各组实验均与进口ｈＥＲＡ／ＢＭｃＡｂ进行比较，结果相符。

Hybridoma secreting specific McAb against hu ER D prepared in our lab was first appraised in ER positive MCF 7 derived stable cell lines by Western blotting. The specific reaction was recognized by immunostaining. Fifty one paraffin embedded sections of breast cancer which were prepared by using ER IHC of hERD McAb and compared with ER DCC yielded an overall concordance rate of 92% between these two technics. The fluorescent mark was positive when it was detected by hERD McAb with indirect immunofluorescence reaction in ER positive MCF 7 cells. Using 3 different paraffin embedded tissues with known ER values as antigen to identify whether or not the self made hERD McAb could be used to detect ER in clinical FCM method. The result showed that ER FCM was accorded with ER IHC. That indicated that hERD McAb can be used to detect ER during procedure of FCM. All the above experiments have been compared with imported hERA/B McAb in good agreement.