Use of streptavidin and biotinylated protein networks in protein signal amplification

• Main Authors: Yu-Wei Chu, 朱峪緯
• Other Authors: James R. Carey
• Format: Others
• Language: en_US
• Published: 2013
• Online Access: http://ndltd.ncl.edu.tw/handle/94277648433967830360

碩士 === 國立高雄大學 === 應用化學系碩士班 === 101 === Enzyme-linked immunosorbent assay (ELISA) is the current method to diagnose presented antigen in human fluids. In recent years, various studies were proceeded base on this platform in order to generate an amplified signal from the antibody-antigen recognition event. Although there are few studies demonstrating a low-level detection limit to detect antigen, the high cost of method is challenged to practice. Therefore a diagnostic device with rapid detection and low cost is needed. We introduce a signal amplification method utilizing the streptavidin-biotin interaction. Since streptavidin is a tetramer for biotin binding, streptavidin can easily form a protein polymer with biotinylated proteins. The method described herein involves streptavidin crosslinking with biotinylated proteins to generate a large detectable protein polymer at the initial antibody-antigen sandwich complex. Specifically, streptavidins and biotinylated proteins are sequentially introduced to generate a. protein polymer possessing a large biotin surface. The biotin surface is then readily available to bind streptavidin labeled signal generating media to generate further signal enhancement of surface antibody-antigen complex. During a mock immunoassay experiment, we have demonstrated a 112 fold decrease in detection limit in comparison with commercial available method.