用自制单克隆抗体建立检测NGAL的胶体金方法

目的:通过免疫、融合、纯化获得了抗中性粒细胞明胶酶相关脂质运载蛋白(NGAL)的特异性单克隆抗体,并建立了胶体金试纸条的检测。方法:本研究筛选得到5株抗NGAL的单克隆抗体,通过配对测试,1B4和2C6能用于双抗夹心胶体金试纸条的制备。以重组hrNGAL为检测对象,对本试纸条的灵敏度、重复性、稳定性进行了全面评价。结果:本试纸条的灵敏度为5ng/ml,重复性和稳定性良好,对临床样本的检测与临床结果一致。结论:本研究建立的NGAL免疫胶体金检测方法,灵敏度高,检测时间短,成本低,为普及临床检测早期肾损伤具有重要意义。

Preparation of anti-NGAL monoclonal antibody and establishment of immunogel gold detection method

Objective: The specific monoclonal antibody against neutrophylase related lipid transport protein (Neutrophils gelatinase,NGAL) was obtained by immunity,fusion purification,and the detection of colloidal gold test notes was established. Methods: Five monoclonal antibodies against NGAL were selected.Through pairing tests,1B4 and 2C6 could be used to prepare double anti-sandwich colloidal gold strips.The sensitivity,repeatability and stability of this test note were evaluated by recombinant hrNGAL. Results: The sensitivity of this test note was 5 ng/ml,and the repeatability and stability were good,the detection of clinical samples was consistent with the clinical results. Conclusion: The method established in this study is of great significance for clinical detection of early renal injury due to its high sensitivity,short detection time and low cost.