| Summary: | 碩士 === 國立成功大學 === 工程科學系碩博士班 === 93 === The design and experiment of this research lie in combining the microchip and nanoparticles to set up a new silver-precipitation immunoassay system. In this study, gold nanoparticle-labeled antibody is coupled with silver enhancement method to provide an alternative of effective and convenient immunoassay. In addition, the immunoassay carried out on the microchip has the advantages of versatile applications, lower sample consumption, shorter detection time and high sensitivity.
Compared with traditional enzyme-linked immunosorbent assay (ELISA), we adopt the immobilization of antigen or antibody on the chip surface, and introduce silver enhancement method to magnify the detection signal generated by the gold nanoparticle. Protein A and immunoglobulin G (IgG) are selected as the model immunoassay to estimate the feasibility and efficiency of the silver-precipitation immunoassay. There are two major formats developed in this study: 1. direct immunoassay (two-layer format), the antigen is immobilized on the chip directly to optimize the assay conditions; 2. sandwich immunoassay (sandwich format), primary and secondary antibodies are used to quantify and qualify antigens.
The results show that the silver-precipitation immunoassay can be measured with optical scanning and electro-signal detection methods. The relationship between concentration and signal is established and the antigen detection limit is 1 ng/mL. The high applicability and biochemical efficiency of this study can provide an alternative for rapid, sensitive and convenient immunoassay.
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