Song Siyu, Lu Yuchao, Li Xueming, Cao Shoupeng, Pei Yuxin, Aastrup Teodor, Pei Zhichao
Shaanxi Key Laboratory of Natural Products & Chemical Biology, College of Chemistry & Pharmacy, Northwest A&F University, Yangling 712100, China.
Attana AB, SE-11419 Stockholm, Sweden.
Polymers (Basel). 2017 Sep 1;9(9):409. doi: 10.3390/polym9090409.
Quartz crystal microbalance (QCM) has been extensively applied in real-time and label-free biomolecular interaction studies. However, the sensitive detection by QCM technology remains challenging, mainly due to the limited surface immobilization capacity. Here, a three-dimensional (3D) carboxymethyl dextran coated gold sensor chip surface was successfully fabricated with dextran of different molecular weight (100, 500 and 2000 kDa, respectively). To evaluate the 3D carboxymethyl dextran surface immobilization capacity, the 3D surface was used for studying antigen⁻antibody interactions on the QCM biosensor. The results showed that the protein immobilization capacity of the 3D carboxymethyl dextran (2000 kDa) surface exceeded more than 4 times the capacity of the 2D carboxyl surface, and 2 times the capacity of the traditional 3D carboxymethyl dextran (500 kDa) surface. Furthermore, the kinetic and affinity properties of antigen⁻antibody interactions were performed. Most notably, the optimized 3D carboxymethyl dextran (2000 kDa) surface could be used for small molecule detection, where the binding of biotinylated oligo (0.67 kDa) reached 8.1 Hz. The results confirmed that a 3D carboxymethyl dextran (2000 kDa) surface can be exploited for sensitive detection of low molecular weight analytes, which have great potential applications for characterizing the interactions between small molecule drugs and proteins.
石英晶体微天平(QCM)已广泛应用于实时、无标记的生物分子相互作用研究。然而,QCM技术的灵敏检测仍然具有挑战性,主要是由于表面固定能力有限。在此,分别用不同分子量(100、500和2000 kDa)的葡聚糖成功制备了三维(3D)羧甲基葡聚糖包被的金传感器芯片表面。为了评估3D羧甲基葡聚糖表面的固定能力,将该3D表面用于研究QCM生物传感器上的抗原-抗体相互作用。结果表明,3D羧甲基葡聚糖(2000 kDa)表面的蛋白质固定能力超过二维羧基表面能力的4倍以上,是传统3D羧甲基葡聚糖(500 kDa)表面能力的2倍。此外,还研究了抗原-抗体相互作用的动力学和亲和力特性。最值得注意的是,优化后的3D羧甲基葡聚糖(2000 kDa)表面可用于小分子检测,其中生物素化寡聚物(0.67 kDa)的结合达到8.1 Hz。结果证实,3D羧甲基葡聚糖(2000 kDa)表面可用于灵敏检测低分子量分析物,这在表征小分子药物与蛋白质之间的相互作用方面具有巨大的潜在应用价值。