An Immunoassay Using Antibody-gold Nanoparticle Conjugate, Silver Enhancement And Flatbed Scanner

摘要

This study reports a convenient immunoassay using antibody-gold nanoparticle (Ab-AuNP) conjugate as a reporter molecule and a flatbed scanner for the optical scanning and measuring of the immuno-reaction signal. The silver enhancement reaction, a signal amplification method in which silver ions are reduced to silver metal, is introduced to magnify the detection signal. The whole framework of the study is based on (1) the direct immunoassay (two-layer format) in which the antigen is directly immobilized on the chip, and (2) the sandwich immunoassay [three-layer format, process similar to enzyme-linked immunosorbent assay (ELISA)], which contains a primary antibody, a secondary antibody (antibody-nanoparticle conjugate) and antigens. The experimental data show that the micro-scale silver precipitation phenomenon is catalyzed by Ab-AuNP conjugates. This reaction can be observed by the naked eye, even at an IgG-AuNPs concentration of 5 pM. The relationships between sample concentration and detection signal are discussed, and the detection limit (sandwich assay) for the sample antigen is 1 ng/mL. Using a flatbed scanner, Ab-AuNP conjugates and a silver enhancement reaction, a new immunoassay is constructed.