Enzymatic amplification combined with nanoparticles-based detection for Microarray
The development of techniques for parallel analysis of biomolecules such as proteins or oligonucleotides is still continuing . Many researchers have always focused their work on achieving an improvement regarding performance and availability. Many significant works represent these efforts. Microarrays are 2D patterns consisting of surface-immobilized molecules of defined molecular species. The goal of our group is to develop a sensitive technique for analyzing signals on protein microarrays by applying the Surface Plasmon Resonance (SPR). We aim to develop a new way for the detection of specific molecular binding, which is based on Rolling Circles Amplification (RCA)  post-signal processing method and optical visualization of nanogold particleslabeled molecules on a micro-structured chip surface (figure 6.1). Besides, the covalent bonding of the RCA prior to the detection of antibodies guaranteed that the surrogate relationship between the analytes and the amplified product was maintained during the investigation. A crucial benefit from applying this new technique in contrast to conventional methods was the expected improved sensitivity and stability. The bleaching effects, which frequently occurred in fluorescence dyes, could not be observed in our nanogold probes. Regarding oligonucleotides modified nanogold particles, regeneration of these detection probes for reusing may be a further advantage. This combination of an easily detectable signal with chip technology should have the potential for high-throughput applications.
KeywordsSurface Plasmon Resonance Bleaching Effect Trisodium Citrate Protein Microarrays Rolling Circle Amplification
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- 4.Hsu, H.-Y. (2003) RCA Combined Nanoparticle-Based Optical Detection Technique for Protein Microarray. Master Thesis, National Taiwan University, Taipei, Taiwan.Google Scholar