物理化学学报 >> 2005, Vol. 21 >> Issue (08): 888-892.doi: 10.3866/PKU.WHXB20050813

研究论文 上一篇    下一篇

蛋白质微阵列SPR实时相位检测

余兴龙; 魏星; 王鼎新; 定翔; 廖玮; 赵新生   

  1. 清华大学精密仪器与机械学系, 精密测试技术及仪器国家重点实验室, 北京 100084; 北京大学化学与分子工程学院化学生物学系, 分子动态与稳态结构国家重点实验室, 北京 100871
  • 收稿日期:2004-12-15 修回日期:2005-03-01 发布日期:2005-08-15
  • 通讯作者: 余兴龙 E-mail:jyxyxl@mail.tsinghua.edu.cn

Real-time SPR Phase Detection for Protein Micro-array

YU Xing-long; WEI Xing; WANG Ding-xin; DING Xiang; LIAO Wei; ZHAO Xin-sheng   

  1. State Key Laboratory of Precision Measurement Technology and Instrument, Department of Precision Instruments and Mechanology, Tsinghua University, Beijing 100084; State Key Laboratory for Structural Chemistry of Unstable and Stable Species, Department of Chemical Biology, College of Chemistry and Molecular Engineering, Peking University, Beijing 100871
  • Received:2004-12-15 Revised:2005-03-01 Published:2005-08-15
  • Contact: YU Xing-long E-mail:jyxyxl@mail.tsinghua.edu.cn

摘要: 将表面等离子体共振(surface plasmon resonance, SPR)与空间相位调制检测结合, 用一束准直的平行光照射传感芯片, 在反射光路中引入渥拉斯顿棱镜, 使光束中的p光和s光产生偏振光干涉形成条纹, 生化反应的有关信息从干涉条纹的相位变化中得出. 选择金膜厚度为30 nm和40 nm的两种芯片, 分别用浓度为50%和20%的酒精-水溶液进行实验, 得到SPR阵列图谱;选择金膜厚度为30 nm, 兔IgG作为阳性对照, 牛血清白蛋白(BSA)作为阴性对照, 与0.2 mL的羊抗兔IgG、1.8 mL水的混合溶液反应, 得到阵列反应图谱. 实验结果表明, 这种方法具有抗干扰能力强, 灵敏度高, 无需标记和可实时检测等优点, 能满足蛋白质芯片检测的要求.

关键词: 蛋白质微阵列, SPR, 相位检测, 偏振光干涉

Abstract: Combining surface plasmon resonance (SPR) and spatial phase modulation, a real-time SPR sensing chip was illuminated with a collimated parallel light beam. A Wollaston prism was introduced into the detection light path, causing the p- and s-polarization components to interfere with each other. The information of biochemical reactions could be obtained by extracting the phase change from the interference patterns. The sensor chips, on which the Au film was 30 nm and 40 nm, experimented with 50% and 20% alcohol solutions respectively to obtain SPR array graph. Subsequently, two proteins, rabbit IgG as positive sample and Bovine Serum Albumin (BSA) as negative control, reacted with the mixed solution of 0.2 mL goat-anti-rabbit IgG sample and 1.8 mL deionized water, using Au film of 30 nm. The reaction diagram was also attained. The experimental results indicated that, with the merits of high noise resistance, high sensitivity, unlabelled and real-time detection, this SPR method could meet the requirement of protein micro-array detection.

Key words: Protein micro-array, SPR, Phase detection, Interference of polarized light