Daneshvar M I, Peralta J M, Casay G A, Narayanan N, Evans L, Patonay G, Strekowski L
Department of Chemistry, Georgia State University, Atlanta 30303, USA.
J Immunol Methods. 1999 Jun 24;226(1-2):119-28. doi: 10.1016/s0022-1759(99)00050-2.
The design, development, and application of a fluorescent fiber-optic immunosensor (FFOI) procedure for the detection of antibody/antigen binding within the near-infrared (NIR) spectral region is reported. The technique was developed through the combined use of fiber-optics, semiconductor laser excitation, fluorescence detection, NIR dye, and immunochemical techniques. The antibody is immobilized on the FFOI's sensing tip and utilized as a recognition component for trace amounts of specific antigen. The FFOI is constructed to utilize antibody sandwich technique. Three individual immunoassays are reported. The first two assays utilize the FFOI and NN382, a commercial NIR dye, for the detection of human immunoglobulin G (IgG). In these assays, goat anti-human IgG antibody (GAHG) is immobilized on the sensitive terminal of the FFOI followed by the exposure of the antibody-coated terminal to human IgG. The probe is then introduced to GAHG labeled with NN382, generating a signal. The third assay utilizes the FFOI for the detection of trace amounts of Legionella pneumophila serogroup 1 (LPS1). In this assay, rabbit anti-LPS1 antibody is immobilized on the sensitive terminal of the FFOI followed by exposure to LPS1. The antigen-coated probe is then treated with monoclonal anti-LPS1 antibody followed by incubation with GAHG labeled with NN382. The assays are optimized to detect the corresponding antigen via the NIR-FFOI. Typical measurements are performed in 10-15 min. A 780-nm semiconductor laser provides the excitation of the immune complex and the resulting emission is detected by a 820-nm silicon photodiode detector. The intensity of the resulting fluorescence is directly proportional to the concentration of the antigen. Solutions of IgG and LPS1 with concentrations as low as 10(-11) M and 0.5 ng/ml, respectively, have been detected with a minimum interference.
报道了一种用于检测近红外（NIR）光谱区域内抗体/抗原结合的荧光光纤免疫传感器（FFOI）方法的设计、开发及应用。该技术是通过光纤、半导体激光激发、荧光检测、近红外染料和免疫化学技术的联合使用而开发的。抗体固定在FFOI的传感尖端上，并用作痕量特异性抗原的识别组件。FFOI采用抗体夹心技术构建。报道了三种独立的免疫测定。前两种测定使用FFOI和商用近红外染料NN382检测人免疫球蛋白G（IgG）。在这些测定中，山羊抗人IgG抗体（GAHG）固定在FFOI的敏感末端，然后将抗体包被的末端暴露于人IgG。然后将探针引入用NN382标记的GAHG中，产生信号。第三种测定使用FFOI检测痕量嗜肺军团菌血清1型（LPS1）。在该测定中，兔抗LPS1抗体固定在FFOI的敏感末端，然后暴露于LPS1。然后用单克隆抗LPS1抗体处理抗原包被的探针，接着与用NN382标记的GAHG孵育。这些测定经过优化，以通过近红外FFOI检测相应抗原。典型测量在10 - 15分钟内完成。780纳米半导体激光激发免疫复合物，产生的发射由820纳米硅光电二极管探测器检测。产生的荧光强度与抗原浓度成正比。分别检测到浓度低至10^(-11) M的IgG溶液和0.5 ng/ml的LPS1溶液，干扰最小。
