摘要
目的建立乳腺癌患者血清HER-2/neu胞外域蛋白(ECD)水平检测方法,为乳腺癌治疗及预后判断提供参考。方法以识别HER-2/neu不同抗原表位的2种抗HER-2/neu单克隆抗体(McAb)为基础,建立可定量检测血清HER-2/neuECD水平的双抗夹心酶联免疫吸附试验法(ELISA)。结果本研究建立的HER-2/neuECD水平检测ELISA所用试剂[包括抗HER-2/neuMcAb5A12包被的酶标板、辣根过氧化物酶(HRP)标记的HER-2/neuMcAb1-2及HRP底物四甲基联苯胺(TMB)溶液]在1年内保持稳定;批内、批间变异系数分别为3·6%±4·2%、6·9%±3·3%;与HER-2ECD同源性很高的人表皮生长因子受体(EGFR/HER-1)ECD不发生交叉反应。自制ELISA试剂盒与BenderMedsystems试剂盒检测人血清HER-2ECD水平的相关性较好(r>0·75)。健康人群(20名)与乳腺癌患者(140例)血清HER-2ECD阳性率之间的差异无统计学意义(P>0·05),但与转移乳腺癌患者(10例)血清HER-2ECD阳性率之间差异有统计学意义(P<0·01)。结论本研究建立的HER-2/neuECD双抗夹心ELISA检测方法特异、稳定、可靠,可定量检测乳腺癌患者血清HER-2/neuECD水平。
Objective To develop an ELISA method to measure HER-2/neu extracellular domain （ECD） levels in sera for predicting the treatment response and prognosis in patients with breast cancer. Methods Anti-HER-2/neu monoclonal antibodies recognizing two different epitopes on HER-2/neu named 5A12 and 1-2 were used as capture antibody and detecting antibody respectively to establish a sandwich enzyme-linked immunosorbent assay （ELISA） ; Horseradish peroxidase activity can be measured quantitatively at 450 nm. Results The ELISA Kit components, including anti-HER-2/neu McAb 5A12 coated on ELISA plates, 1-2 labeled with HRP and the HRP substrate solution （0.01% 3,3＇,5,5＇- tetramethylbenzidine,TMB） were stable for one year. Intra-and inter-run imprecision （CV） were 3. 6% ± 4. 2% and 6. 9% ± 3. 3%, respectively. No cross-reaction with human epidermal growth factor receptor （EGFR/HER-1） ECD, which has extensive homology with HER-2 ECD, was found. There was good correlation （r 〉 0. 75 ）for HER-2 ECD levels in human sera between Bender Medsystems and our ELISA;The positive rate of HER-2 ECD in sera of healthy subjects was no difference to that of breast cancer patients （ P 〉 0. 05 ）, but significant differenceto that of metastatic breast cancer patients （ P 〈 0. 01 ）. Conclusion The ELISA kit for serum HER-2/neu ECD levels from breast cancer patients was specific, reliable and could be quantified.
出处
《中华检验医学杂志》
CAS
CSCD
北大核心
2006年第4期310-313,共4页
Chinese Journal of Laboratory Medicine
基金
首都发展基金科研项目资助课题(2002-2023)
