化学学报 ›› 2009, Vol. 67 ›› Issue (17): 2013-2018. 上一篇    下一篇

研究论文

痕量铜蓝蛋白的免疫纳米金催化-氧化亚铜微粒共振散射光谱分析

刘庆业a 韦丽丽a 梁爱惠*,a 蒋治良*,a 王素梅b

  

  1. (a广西师范大学环境与资源学院 桂林 541004)
    (b桂林工学院材料与化学工程系 有色金属及材料加工新技术教育部重点实验室 桂林 541004)

  • 投稿日期:2008-06-20 修回日期:2009-05-05 发布日期:2009-09-14
  • 通讯作者: 梁爱惠

Resonance Scattering Spectral Assay of Trace Ceruloplasmin Using Immunonanogold-fehling Reagent-glucose Catalytic Reaction

Liu, Qingyea Wei, Lilia Liang, Aihui*,a Jiang, Zhiliang*,a Wang, Sumeib   

  1. (a School of Environment and Resource, Guangxi Normal University, Guilin 541004)
    (b Key Laboratory of New Processing Technology for Nonferrous Metals and Materials of Education Ministry,
    Department of Material and Chemical Engineering, Guilin University of Technology, Guilin 541004)
  • Received:2008-06-20 Revised:2009-05-05 Published:2009-09-14
  • Contact: Liang, Aihui

用15 nm的纳米金标记羊抗人铜蓝蛋白抗体(GCP)可获得铜蓝蛋白(CP)纳米金探针(AuGCP). 在pH 7.8柠檬酸-磷酸氢二钠缓冲溶液中, CP与AuGCP发生特异性结合生成胶体金免疫复合物. 离心分离后, 离心液中的AuGCP可作为酒石酸铜(C4H4O6Cu)-葡萄糖反应体系的催化剂, 生成的Cu2O微粒在620 nm处有一共振散射峰. 在选定条件下, 620 nm处共振散射信号降低值△I620 nm与铜蓝蛋白浓度cCP在0.18~45 ng/mL范围内存在良好线性关系, 回归方程为
ΔI620 nm=2.27cCP+5.05, 相关系数为0.9940, 检出限为0.14 ng/mL. 该法用于人血清中铜蓝蛋白的检测, 结果满意.

关键词: 铜蓝蛋白, 免疫纳米金催化, 氧化亚铜微粒, 葡萄糖, 共振散射光谱法

15 nm nanogold was used to label goat anti-human ceruloplasmin (GCP) to obtain an immunonaogold resonance scattering (RS) probe (AuGCP) for ceruloplasmin. In the pH 7.8 citric acid-Na2HPO4 buffer solution, the immunoreaction between nanogold-labeled goat anti-human ceruloplasmin (AuGCP) and CP took place to form a nanogold-labeled immune complex. After centrifugation, the AuGCP in the centrifugal solutions could be used as catalyst for the reaction between Fehling reagent and glucose. The product of Cu2O particles has the strongest resonance scattering (RS) peak at 620 nm. Under the selected conditions, the decreased intensity ΔI620 nm is proportional to CP concentration (cCP) from 0.18 to 45 ng/mL, with a regression equation of ΔI620 nm=2.27cCP+5.05, with a correlation coefficient of 0.9940, and a detection limit of 0.14 ng/mL CP. This simple and sensitive RS assay was applied to determination of CP in human plasma with satisfactory results.

Key words: ceruloplasmin, immunonanogold catalysis, Cu2O particle, glucose, resonance scattering spectral assay