SPR (Surface Plasmon Resonance) is an emerging optical biosensor which can monitor the processes at metal interface in real time without labeling requirements.Instrument SPR-2004 can sense two areas in one channel and...SPR (Surface Plasmon Resonance) is an emerging optical biosensor which can monitor the processes at metal interface in real time without labeling requirements.Instrument SPR-2004 can sense two areas in one channel and get two signals at same time in one test.One signal is the graph of the reaction;the other is the graph of reference.SPR biochip is modified with dextran,and only reaction area can be used to immobilize the protein.Two methods of antibody immobilization on chip were tested.One was to immobilize directly,the other was to immobilize protein A firstly and then the antibody was caught by immobilized protein A.The latter was chosen as suitable for retention of the native binding ability with virus of antibody.The latter chip was used to detect SARS-Cov.The signal of detection reached to 60 units within 40 min.It was 55 units even after reference.展开更多
Cardiac troponin I(cTnI) was separated and purified from human left ventricular tissue by affinity chromatographic method and used to immunize Balb/c mice by intraperitoneal injection and four hybridoma cell lines, wh...Cardiac troponin I(cTnI) was separated and purified from human left ventricular tissue by affinity chromatographic method and used to immunize Balb/c mice by intraperitoneal injection and four hybridoma cell lines, which secreted monoclonal antibody(mAb) against human cTnI, were obtained by cell fusion, identification and cloning twice. Three mAbs(9F5, 2F11, 8C12) were produced from the ascites of Balb/c mice injected intraperitoneally the hybridoma cells and characterized by means of a surface plasmon resonance(SPR) biosensor. An optimal and specific sensing membrane for troponin I was prepared with staphylococcal protein A(SPA) as the intermediate layer and mAb against human cTnI as the capture antibody. On the basis of the sensing membrane, two modes of operation of the SPR biosensor were developed, i.e ., a direct detection of antigen antibody affinity and a sandwich assay. In the sandwich assay detection mode, the mAbs competition was measured by monitoring whether the secondary antibody had been attached to the cTnI already captured by the first antibody on the sensor surface. The SPR biosensor was shown to be able to directly detect the antigen antibody affinity and the order of the affinity was found to be 9F5>2F11>8C12. In the sandwich detection mode, it was found that the different epitopes on the cTnI molecules were recognized by the three mAbs respectively, but the asymmetrical competition was shown between 2F11 and 8C12 and no competition was found between 9F5 and 2F11 or 8c12. Based on these results, a double monoclonal sandwich immunoassay for cTnI was developed by using the optimal antibody pair of 9F5 and 2F11 and the SPR biosensor with SPA substrate membrane, which showed an excellent sensitivity of 0.8 μg/L for both the buffer and the serum samples compared with the direct detection of cTnI for the buffer with the lowest detection limit of 4 μg/L and conventional ELISA with the sensitivity of 1.9 μg/L.展开更多
表面等离子体谐振(surface p lasm on resonance,SPR)生物传感器是一种基于物理光学原理的新型生化分析系统。与传统的相互作用分析技术相比较,它具有实时监控、无需标记、耗样量极少等特点,在研究分子间相互作用、药物残留检验、临床...表面等离子体谐振(surface p lasm on resonance,SPR)生物传感器是一种基于物理光学原理的新型生化分析系统。与传统的相互作用分析技术相比较,它具有实时监控、无需标记、耗样量极少等特点,在研究分子间相互作用、药物残留检验、临床疾病诊断以及畜牧养殖业等诸多方面都具有应用价值。作者对近几年国内和国际上SPR生物传感器的应用进展和发展趋势进行了简要综述。展开更多
基金This work was supported by the National Natural Science Foundation of China （No. 60341005 and No. 60427001）.
文摘SPR (Surface Plasmon Resonance) is an emerging optical biosensor which can monitor the processes at metal interface in real time without labeling requirements.Instrument SPR-2004 can sense two areas in one channel and get two signals at same time in one test.One signal is the graph of the reaction;the other is the graph of reference.SPR biochip is modified with dextran,and only reaction area can be used to immobilize the protein.Two methods of antibody immobilization on chip were tested.One was to immobilize directly,the other was to immobilize protein A firstly and then the antibody was caught by immobilized protein A.The latter was chosen as suitable for retention of the native binding ability with virus of antibody.The latter chip was used to detect SARS-Cov.The signal of detection reached to 60 units within 40 min.It was 55 units even after reference.
基金Supported by National Natural Science Fundation of China(Project No.2 9875 0 10 ) and the Health Department of JilinProvince(Project No.980 4 7) ,China
文摘Cardiac troponin I(cTnI) was separated and purified from human left ventricular tissue by affinity chromatographic method and used to immunize Balb/c mice by intraperitoneal injection and four hybridoma cell lines, which secreted monoclonal antibody(mAb) against human cTnI, were obtained by cell fusion, identification and cloning twice. Three mAbs(9F5, 2F11, 8C12) were produced from the ascites of Balb/c mice injected intraperitoneally the hybridoma cells and characterized by means of a surface plasmon resonance(SPR) biosensor. An optimal and specific sensing membrane for troponin I was prepared with staphylococcal protein A(SPA) as the intermediate layer and mAb against human cTnI as the capture antibody. On the basis of the sensing membrane, two modes of operation of the SPR biosensor were developed, i.e ., a direct detection of antigen antibody affinity and a sandwich assay. In the sandwich assay detection mode, the mAbs competition was measured by monitoring whether the secondary antibody had been attached to the cTnI already captured by the first antibody on the sensor surface. The SPR biosensor was shown to be able to directly detect the antigen antibody affinity and the order of the affinity was found to be 9F5>2F11>8C12. In the sandwich detection mode, it was found that the different epitopes on the cTnI molecules were recognized by the three mAbs respectively, but the asymmetrical competition was shown between 2F11 and 8C12 and no competition was found between 9F5 and 2F11 or 8c12. Based on these results, a double monoclonal sandwich immunoassay for cTnI was developed by using the optimal antibody pair of 9F5 and 2F11 and the SPR biosensor with SPA substrate membrane, which showed an excellent sensitivity of 0.8 μg/L for both the buffer and the serum samples compared with the direct detection of cTnI for the buffer with the lowest detection limit of 4 μg/L and conventional ELISA with the sensitivity of 1.9 μg/L.
文摘表面等离子体谐振(surface p lasm on resonance,SPR)生物传感器是一种基于物理光学原理的新型生化分析系统。与传统的相互作用分析技术相比较,它具有实时监控、无需标记、耗样量极少等特点,在研究分子间相互作用、药物残留检验、临床疾病诊断以及畜牧养殖业等诸多方面都具有应用价值。作者对近几年国内和国际上SPR生物传感器的应用进展和发展趋势进行了简要综述。
