黄曲霉毒素的免疫检测方法选读.ppt

The Immunology-based Methods for Aflatoxins Detection Nutrition and Food Hygiene Ⅰ Classification of immunoassay 1. indirect immunoassay 2. sandwich immunoassay Ⅱ Enzyme-linked immunosorbent assays Enzyme immunoassay is a good alternative to chromatography, because it allows an analysis of massive number of samples, easily automatized and does not require time-consuming procedures and sophisticated equipment. Presently enzyme-linked immunosorbent assay (ELISA) is widely accepted as the‘‘gold standard’’ screening method. In ELISA , horseradish peroxidase (HRP) is commonly used as a label of immunoreagents. Multiple detection methods for the enzyme activity of peroxidase-labeled immunoreagents are applied, including colorimetry, fluorimetry and chemiluminescence (CL). CL detection is markedly more sensitive than other methods. Ⅲ Gold immunochromatography assay The nano-gold colloidal is negatively charged, it can easily bind with the sulfhydryl in protein. So it can combine with the antigens and antibodies. Aflatoxin if present in the sample extract interacts with gold nanoparticles conjugated anti-aflatoxin antibodies at the base of the stick. Both bound and unbound antibodies move along the stick membrane, passing a test line composed of immobilised free antibodies to form a visible line indicating a level of aflatoxin contamination. Ⅳ Biosensors Biosensors were developed for the desire of multiple analyses, they are can be used for simultaneous analysis of multiplesamples or simultaneous analyses of multiple target analytes. Biosensors have been developing to take the advantages of nanoscale materials. Immobilization of antibodies on solid–liquid inter-faces is a critical issue in the development of such systems.The sensitivity of immunosensor depends on the surface concentrations of immobilized antibodies, their remaining antigen binding ability and orientation of these molecules at the interface. According to a signal-transfer process, the immunoassays