Detection Of Horseradish Peroxidase With Resonance Light Scattering And Functionalized Carbon Nanotube And It's Applications In Bioassay

Enzyme is a protein which has catalytic activity and the enzyme analysis has great influence in the field of life science research. But the traditional methods to detect enzyme, such as the radioactive label and the enzyme label, generally have complex operations that has limited the advanced research in enzyme in some degree. With the development of life science research, how to establish rapid, simple and highly effective methods to detect enzyme has been becoming a hot topic. HRP (Horseradish Peroxidase) is one of the enzymes which are most widely used. CNT (Carbon Nanotube) is becoming a hot topic for its unique physical and chemical properties after the 90th. Based on above, this paper is divided into two parts to research the HRP and CNT and prospect on the application in the field of the environmental microbiological immunoassay.Part one:Determination of HRP based on the oxidation reaction of phenol with H2O2 by using resonance light scattering technique.A new assay of HRP is proposed by using of resonance light scattering (RLS). The oxidation reaction of phenol with hydrogen peroxide (H2O2) catalyzed by HRP results in a great enhancement of RLS intensity in the wavelength range from 200 nm to 700 nm characterized by the RLS peak around 313 nm. The enhanced intensity of RLS at 313 nm is proportional to the concentration of HRP in the range of 0.05～5.0×10-7 g/mL and the detection limit is 1.1×10-10 g/mL. Therefore, a simple and sensitive method for HRP determination is established.Part two:Chemical modified CNT and DNA molecular functional CNT.We developed applications of CNT in the detection of DNA by using high sensitive fluorescence assay. The amount of signal was proportional to the concentration of target oligonucleotide. At the same time, we lay a good foundation for the application of PFP (cationic conjugated polymer) in process of hybridization reaction. If we design other suitable probes and complementary sequences, this assay could be used for other DNA hybridization and SNP analysis.