基于近红外上转换纳米探针的固相免疫检测研究

以氯化物为原料通过溶剂热法合成了尺寸均匀的NaYF4∶Yb3+,Tm3+上转换纳米粒子(UCNPs), 利用聚丙烯酸(PAA)取代UCNPs表面的油酸配体, 引入羧基和人IgG的氨基共价结合(H端), 再用蛋白G(protein G)作“桥”将兔抗羊IgG(r-a-g IgG)修饰在硅片表面(R端), 以消除由于硅片“刚性”表面与r-a-g IgG分子直接偶联而导致其结构或构象的变化, 从而实现硅片表面与r-a-g IgG的“柔性”生物偶联。利用抗原抗体的特异性免疫反应, 将H端和R端通过羊抗人IgG(g-a-h IgG)结合, 建立了简单高效和快速灵敏检测溶液中g-a-h IgG的新型免疫分析方法。实验结果表明: g-a-h IgG在5~400 nmol/L浓度范围内与上转换荧光强度具有良好的线性关系, 该传感器检测限为1.82 nmol/L, 并表现出优秀的检测特异性。本项研究为临床各种重大疾病的免疫分析诊断提供了一种宽线性范围和高特异性的新型免疫方法和平台。

Abstract

Uniformed NaYF4∶Yb3+,Tm3+ nanoparticles were synthesized through solvent thermal method in chloride. PAA-based ligand replace strategies were carried out to introduce free carboxylic acid groups which allowed further conjugation with NH2-containing human IgG. The silicon wafer was modified with rabbit anti-goat IgG by using protein G as “bridge”, which contributed to eliminate structural or conformational changes of IgG. Owing to the specific antibody recognition to the antigenic, a simple, rapid, efficient and sensitive immunoassay method was developed. The experimental results demonstrate that there is a very good linear correlation between the upconversion luminescence intensity and the amount of goat anti-human IgG in the range of 5~400 nmol/L. The detection limit is as low as 1.82 nmol/L. Our developed detection system shows excellent detection specificity and sensitivity, which provides a new detection method and platform with wide linear range and high sensitivity for diagnosis of major clinical diseases in the future.

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周进, 张美玲, 张俐, 李翠霞, 赵慧颖, 张友林, 夏安东, 孔祥贵. 基于近红外上转换纳米探针的固相免疫检测研究[J]. 发光学报, 2018, 39(8): 1059. ZHOU Jin, ZHANG Mei-ling, ZHANG Li, LI Cui-xia, ZHAO Hui-ying, ZHANG You-lin, XIA An-dong, KONG Xiang-gui. Studies of Solid Phase Immunoassay Based on Near Infrared Upconversion Nanoprobe[J]. Chinese Journal of Luminescence, 2018, 39(8): 1059.

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