To overcome these issues, microfluidic immunoassay systems have been thenthereby introduced because of their various advantages, including high throughput, high-efficiency, low-cost Inhibitors,Modulators,Libraries and minimized consumption of samples and reagents [5]. After the development of soft lithography techniques using poly(dimethylsiloxane) (PDMS), PDMS has become the most popular microfluidic device materials and offers several advantages such as easy handling, good sealing properties and high optical transparency [6]. However, the poor chemical stability in different types of organic solvents, difficulty in surface modification and mass production have limited the use of PDMS in the various applications [7].Recently, because of the material issues, some researchers have been attempted to use plastic materials as an alternative solution.
Among the various Inhibitors,Modulators,Libraries types of polymers, cyclic olefin copolymer (COC) is one of the most popular polymeric materials in the fabrication of Inhibitors,Modulators,Libraries microfluidic chips. COC is a well-known polymeric material with various advantages, Inhibitors,Modulators,Libraries including high clarity and light transmission, excellent mechanical properties and great biocompatibility [8].Furthermore, effective immobilization of proteins is essential and important in microfluidic chips to be used as immunosensors. Several methods to immobilize antibodies on the sensor chip surface have been developed, including physical adsorption, covalent binding, and specific interaction between avidin and biotin [9,10]. However, these previous methods have limitations in terms of denaturization, extra chemical modification and random orientation.
In order to overcome these issues, Brown et al. and Park et al. developed specific gold-binding polypeptide (GBP) that endows the orientation of proteins in their functional state [11,12]. GBP Anacetrapib shows a strong binding affinity to the gold surface without any surface modifications [13�C15]. Therefore, GBP-fusion proteins could be selectively and functionally immobilized onto the gold surface.In this study, we carefully designed microfluidic devices, and the surface of a detection chamber was coated with gold for the direct assembly of proteins. A microfluidic-based immunosensor to detect human H1N1 influenza was developed into a low-cost immunosensor based on the exploration of fluorescence signals.
The detection of a specific antibody among serological assays in blood samples was performed in the microfluidic biosensor chip by immunoreactions between the GBP-recombinant influenza hemagglutinin antigen (GBP-H1a) fusion protein and its specific antibody (Ab). The GBP-H1a fusion protein as a bioreceptor and the fluorescence-labeled Ab as a marker were used to provide an excellent always find useful information detection signal. In addition, the chip fabrication and sensing characteristics are reported in detail.2.?Experimental Section2.1. Reagents and MaterialsCOC was purchased from TOPAS Advanced Polymers (Frankfurt-H?chst, Germany).