The Preparation And Optimization Of Phycobiliprotein Fluorescence Probes

Immunoassay technology has been widely used in biomedical, environmental monitoring, food inspection and other fields,especially in recent years, a new immunoindicator technology is developing by leaps and bounds. The immunoindicator was classified into radioactive immunoindicator and nonradioactive immunoindicator by marker type. The inradioactive immunoindicator is more widely used because of its remarkable characteristics, such as global green, safe and noncontaminating, especially the fluorescent immunoindicator marked by phycobiliproteins which is one of the most sensitive methods.In this research, we constructed a fusion protein consisting SA and CpcB on the basis of protein CpcB.By reconstitution CpcB with phycobilin, the fluorescence can be detected by observing the light emission. But the fusion protein directly got from the E.coli is not good, so we get it by reconstitution out of the E.coli. SA has the property to effectively binding to biotin, that is something like what we called the BAS system. The fusion protein could be applied to immunoassay, as a new alternative reagent.The fluorescence activity of the fusion protein was tested by reconstitution in vivo and the immunocompetence was tested by immunoreaction. The results show that the fusion protein retained the bifunctional effects of the two proteins. Then by using PEB-CpcB-SA as a second antibody, and testing its fluorescence, we believed it can be used for fluoroimmunassay.