利用表面等离子体共振技术检测人血清中的炭疽保护性抗原，一种炭疽特异性毒素。
Detection of protective antigen, an anthrax specific toxin in human serum by using surface plasmon resonance.
机构信息
Biotechnology Division, Defence Research & Development Establishment, Gwalior-474 002, India.
出版信息
Diagn Microbiol Infect Dis. 2013 Sep;77(1):14-9. doi: 10.1016/j.diagmicrobio.2013.05.006. Epub 2013 Jun 15.
In this study, surface plasmon resonance (SPR) technology was used for the sensitive detection of protective antigen (PA), an anthrax specific toxin in spiked human serum samples. A monoclonal antibody raised against Bacillus anthracis PA was immobilized on carboxymethyldextran-modified gold chip, and its interaction with PA was characterized in situ by SPR. By using kinetic evaluation software, KD (equilibrium constant) and Bmax (maximum binding capacity of analyte) were found to be 20 fM and 18.74 m°, respectively. The change in Gibb's free energy (∆G= -78.04 kJ/mol) confirmed the spontaneous interaction between antigen and antibody. The assay could detect 1 pg/mL purified PA. In PA-spiked human serum samples, 10 pg/mL of PA could be detected. Presence of PA in blood samples serves as an important early diagnostic marker for B. anthracis infections. Thus, SPR test can be a sensitive assay for detection of anthrax at early stages of infection.
在这项研究中，表面等离子体共振（SPR）技术被用于检测炭疽特异性毒素保护性抗原（PA）在加标人血清样本中的灵敏检测。针对炭疽杆菌 PA 产生的单克隆抗体被固定在羧甲基葡聚糖修饰的金芯片上，并用 SPR 对其与 PA 的相互作用进行了原位表征。通过使用动力学评估软件，发现 KD（平衡常数）和 Bmax（分析物的最大结合容量）分别为 20 fM 和 18.74 m°。吉布斯自由能的变化（∆G= -78.04 kJ/mol）证实了抗原和抗体之间的自发相互作用。该测定法可以检测 1 pg/mL 的纯化 PA。在加标人血清样本中，可以检测到 10 pg/mL 的 PA。PA 在血液样本中的存在是炭疽杆菌感染的重要早期诊断标志物。因此，SPR 试验可以作为一种灵敏的检测方法，用于在感染的早期阶段检测炭疽。
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