Recombinant Green Electrochemiluminescent Proteins And Their Immunoassay Applications

Novel analytical apparatuses and methods with high sensitivity,wide versatility and user-friendliness were urgently required in the food safety,environmental protection and clinical diagnosis.Electrochemiluminescence with high sensitivity,good controllability,fast analysis speed and wide range of applications in detection applications,has been widely used in food analysis and environmental testing,biochemical reagent detection and immunoassay analysis.Many.materials have electrochemiluminescence,and finding more efficient luminophores or modifying luminophores for biomolecular labeling has received extensive interests.Among them,as an electrochemiluminescent device,porphyrin has unique luminescent properties and the stability and efficiency of electrochemiluminescence of the porphyrin luminescent material can be improved by assembling the porphyrin with DNA,nanomaterials or suitable biological proteins.In this study,we focus on porphyrin-based electroluminescent materials for experiments.First,a porphyrin compounds with better chemiluminescence propertie was seleted.Cytochrome C,myoglobin and hemoglobin were used as raw material which were separated to prepare apo-cytochrome C,apo-myoglobin and apo-hemoglobin in acidic conditions.Then the apoprotein is combined with porphyrin to prepare a reconstituted protein.The results of ultraviolet spectroscopy,fluorescence spectroscopy and electrochemiluminescence detection show that compared with porphyrin monomer,reconstituted hemoglobin and reconstituted myoglobin exhibit chemluminescence with more stability and activity.The early cancer marker-vascular upper epidermal growth factor detection sensor built by apo-myoglobin was proved to have high detection sensitivity,good selectivity and stability,and has great value in practical applications.