高等学校化学学报

高等学校化学学报 ›› 2001, Vol. 22 ›› Issue (10): 1645.

• 研究论文 • 上一篇    下一篇

固体基质室温磷光免疫分析法测定人IgG

刘佳铭, 付艳, 余冰宾, 李隆弟   

  1. 清华大学化学系, 北京 100084
  • 收稿日期:2000-09-26 出版日期:2001-10-24 发布日期:2001-10-24
  • 通讯作者: 李隆弟(1939年出生),男,教授,主要从事光致发光和分子识别研究.*福建漳州师范学院化学系访问学者.
  • 基金资助:

    国家自然科学基金(批准号:29775013)资助

Determination of IgG in Human Serum by Solid Substrate Room Temperature Phosphorescence-Immunoassay

LIU Jia-Ming, FU Yan, YU Bing-Bin, LI Long-Di   

  1. Department of Chemistry, Tsinghua University, Beijing 100084, China
  • Received:2000-09-26 Online:2001-10-24 Published:2001-10-24

PDF (PC)

被引次数

19

摘要： 在聚酰胺膜(PM)等固体基质上,人免疫球蛋白(IgG)能与异硫氰酸荧光素(FITC)标记的羊抗人抗体(GAHAb*)发生定量的特异性免疫反应,并保持了FITC优良的室温磷光特性,据此建立了固体基质室温磷光免疫分析(SS-RTP-IA)测定人血清中人IgG的新方法.在0.4μL(取样浓度为1.56～50ng/mL)点样体积内,人IgG的含量在0.624～20pg范围内,与室温磷光强度呈良好的线性关系,工作曲线的回归方程ΔIp=31.4+46.2lg[c(Ag)/pg],n=6.相关系数r=0.999,检出限0.32pg.用于正常人血清中人IgG含量的测定,加标回收率为96%～104%,结果满意.

关键词: 固体基质室温磷光免疫分析法, 人IgG, 羊抗人抗体

Abstract: On many kinds of solid substrate, such as polyamide membrane, cellulose acetate membrane etc., the specific immunological reaction between the human immunoglobulin G(IgG) and the goat anti human antibody labeled with isothiocyanate fluorescein(GAHAb *) can be carried into practice quantitatively. The antigen antibody conjugate remained excellent room temperature phosphorescence(RTP) properties of isothiocyanate fluorescein. The conditions for immunological reaction and RTPmeasurement were optimized. Aconvenient solid substrate room temperature phosphorescence immunoassay(SS-RTP-IA) method for determination of the IgGhas been developed. The linear dynamic range of the method is 0.624-20 pg/spot(0.4μL) and the calculated detection limit by the methodology of IUPACis 0.32 pg/spot. The regression equation is ΔIp=31.4+46.2 lg[ c (Ag)/pg], correlation coefficient r=0.999, n=6. The method was used for the determination of Ig Gin normal human serum with good satisfactory results. The analytical recoveries are 96%-104% for seven replicates and the relative standard deviations are 2.6%-2.9%.

Key words: Solid substrate room temperature phosphorescence-immunoassay, Human IgG, GAHAb

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