Ben-Dov I, Willner I, Zisman E
Institute of Chemistry, Hebrew University of Jerusalem, Israel.
Anal Chem. 1997 Sep 1;69(17):3506-12. doi: 10.1021/ac970216s.
The assembly of a biosensor for Chlamydia trachomatis based on the microgravimetric quartz crystal microbalance (QCM) analysis of the bacteria association to an antibody-functionalized electrode is described. The sensing interfaces consist of a primary cystamine monolayer assembled onto Au electrodes associated with the quartz crystal. The monolayer is further modified with sulfosuccinylimidyl 4-(p-maleimidophenyl)butyrate (sulfo-SMPB) and the goat IgG-anti-mouse IgG Fc-specific Ab or the fragmented F(ab')2 anti-mouse IgG Ab that act as sublayers for the association of the sensor-active anti-C. trachomatis LPS-Ab. Bacteria in the concentration range from 260 ng.mL-1 to 7.8 micrograms.mL-1 are sensed by the functionalized crystals. The association of C. trachomatis to the sensing interface can be confirmed and amplified via interaction of the crystal with various anti-C. trachomatis antibodies. Urine-pretreated functionalized quartz crystals are applied in the analysis of C. trachomatis in urine samples. The sensitivity limits of the electrodes for sensing the bacteria in urine samples corresponds to approximately 260 ng.mL-1. The functionalized crystals assembled via association of anti-C. trachomatis LPS-Ab to the fragmented F(ab')2 anti-mouse IgG Ab reveal long-term stability upon storage at 4 degrees C.
本文描述了一种基于微重力石英晶体微天平(QCM)分析细菌与抗体功能化电极结合情况的沙眼衣原体生物传感器的组装。传感界面由组装在与石英晶体相连的金电极上的初级胱胺单层组成。该单层进一步用4-(对马来酰亚胺苯基)丁酸磺基琥珀酰亚胺酯(磺基-SMPB)以及山羊IgG-抗小鼠IgG Fc特异性抗体或片段化的F(ab')2抗小鼠IgG抗体进行修饰,这些抗体作为传感活性抗沙眼衣原体脂多糖抗体结合的子层。功能化晶体可检测浓度范围为260 ng.mL-1至7.8微克.mL-1的细菌。沙眼衣原体与传感界面的结合可通过晶体与各种抗沙眼衣原体抗体的相互作用得到确认和放大。尿液预处理的功能化石英晶体用于尿液样本中沙眼衣原体的分析。电极检测尿液样本中细菌的灵敏度极限约为260 ng.mL-1。通过抗沙眼衣原体脂多糖抗体与片段化的F(ab')2抗小鼠IgG抗体结合组装的功能化晶体在4℃储存时显示出长期稳定性。