上转换纳米粒子NaYF4:Yb,Er的免疫荧光淬灭法测定中性粒细胞明胶酶相关载脂蛋白
Determination of Neutrophil Gelatinase-Associated Lipocalin Based on Immune Fluorescence Quenching of NaYF4:Yb, Er Upconversion Nanophosphors
摘 要
采用上转换纳米粒子（UCP）NaYF4：Yb，Er和量子点（QDs）CdTe分别标记中性粒细胞明胶酶相关载脂蛋白（NGAL）的两株抗体，标记抗体与NGAL通过免疫反应形成双抗夹心体，无须洗涤分离直接测定。结果表明：UCP在540 nm处的荧光可有效淬灭。NGAL的质量浓度在40~750 μg·L-1内与UCP的光强比呈线性关系，检出限（3s/k）为23.3 μg·L-1。以空白样品为基体进行加标回收试验，所得回收率为94.6%~108%，测定值的相对标准偏差（n=6）为5.5%~9.1%。
荧光淬灭
中性粒细胞明胶酶相关载脂蛋白
免疫测定
upconversion nanophosphor
fluorescence quenching
neutrophil gelatinase-associated lipocalin
immunoassay
Abstract
Two antibodies of neutrophil gelatinase-associated lipocalin (NGAL) were labeled by NaYF4:Yb,Er upconversion nanophosphors (UCP) and CdTe quantum dots (QDs), respectively. The labelled-antibodies reacted with NGAL forming double anti-sandwich body by immune reaction and NGAL was determined without any washing and separation operation. It was found that fluorescence of UCP at 540 nm could significantly quenched. Linear relationship between values of fluorescence intensity ratio of UCP and mass concentration of NGAL was obtained in the range of 40-750 μg · L-1, with detection limit (3s/k) of 23.3 μg·L-1. On the base of blank sample, test for recovery was made by standard addition method. Values of recovery found were in the range of 94.6%-108%, with RSD's (n=6) in the range of 5.5%-9.1%.
中图分类号 O657.3 DOI 10.11973/lhjy-hx201802001
所属栏目 试验与研究
基金项目 国家自然科学基金（21176124）
收稿日期 2017/2/28
修改稿日期
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备注邵祥明(1991-),男,江苏淮安人,硕士研究生,主要从事免疫分析研究工作
引用该论文: SHAO Xiangming,FENG Hui,ZHOU Yang,HAN Yuwang,ZHOU Taoyu,ZHANG Hongman. Determination of Neutrophil Gelatinase-Associated Lipocalin Based on Immune Fluorescence Quenching of NaYF4:Yb, Er Upconversion Nanophosphors[J]. Physical Testing and Chemical Analysis part B:Chemical Analysis, 2018, 54(2): 125～129
邵祥明,冯慧,周洋,韩毓旺,周桃玉,张红漫. 上转换纳米粒子NaYF4:Yb,Er的免疫荧光淬灭法测定中性粒细胞明胶酶相关载脂蛋白[J]. 理化检验－化学分册, 2018, 54(2): 125～129
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【2】HIRSCH R, DENT C, PFRIEM H, et al. NGAL is an early predictive biomarker of contrast-induced nephropathy in children[J]. Pediatric Nephrology, 2007,22(12):2089-2095.
【3】齐家龙,邵佳,彭宽,等.中性粒细胞明胶酶相关脂质运载蛋白的单克隆抗体制备及化学发光免疫定量检测试剂研究[J]. 中国生化药物杂志, 2015,35(4):5-9.
【4】赵元明,李新建,马生聘,等.联合检测尿中NGAL、mAlb、α1-MG在糖尿病肾病中的早期诊断价值[J].中华全科医学, 2015,13(10):1678-1680.
【5】BACHORZEWSKA-GAJEWSKA H, MALYSZKO J, SITNIEWSKA E, et al. Neutrophil-gelatinase-associated lipocalin and renal function after percutaneous coronary interventions[J]. American Journal of Nephrology, 2006,26(3):287-292.
【6】张龙山,李倩.人尿液NGAL蛋白免疫比浊检测方法的建立和临床检测意义[J].临床和实验医学杂志, 2016,15(11):1135-1136.
【7】HAASE M, SCHÄFER H. Upconverting nanoparticles[J]. Angewandte Chemie International Edition, 2011,50(26):5808-5829.
【8】XU C T, ZHAN Q Q, LIU H, et al. Upconverting nanoparticles for pre-clinical diffuse optical imaging, microscopy and sensing:Current trends and future challenges[J]. Laser & Photonics Reviews, 2013,7(5):663-697.
【9】LIU Q, FENG W, YANG T, et al. Upconversion luminescence imaging of cells and small animals[J]. Nature Protocol, 2013,8(10):2033-2044.
【10】刘丽萍,周皓东,汤杰,等.基于CdTe量子点测定中性粒细胞明胶酶相关载脂蛋白的免疫荧光层析法[J].理化检验-化学分册, 2016,52(12):1365-1369.
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