The Application In Detection Of Biomolecules By Coupling Technique

Metalloprotein is a kind of protein with metal ion cofactor.It exerts important functions in cells.As an important metalloprotein, metallothionein (MT) is characterized by low molecular weight,a high cysteine and metal content and possesses a highly conserved structure.MT have been purported to play a important role in regulation of essential metals,detoxificaiton of heavy metals and scavenging of free radicals.To detect MT rapidly and accurately,we have developed a method which UV-vis spectrophotometry combined in tandem with voltammetry for MT investigation.MT sample and the electrophilic disulphide DTDP (4,4’-dithiodipyridine) were first introduced to the UV-vis detector by the carrier solution for spectrophotometric thiol assays and then delivered into the electrochemical flow cell, metal ion was detected. With this coupled technique, uncertainties that cannot be addressed by the individual techniques are largely clarified.Such a hyphenated system is amenable to the accurate and convenient determination of MT and should be useful for physiological studies.MicroRNA play an important role in life activity, especially its effect in tumor occur and deterioration.In order to detect microRNA at low detection level in human-being,we coupled flow-injection with surface plasmon resonance.Because SA-HRP can specific binding to biotin-RNA,the HRP-biocatalyzed oxidation of4-chloro-l-naphthol (4-CN) in the presence of H2O2,the subsequent precipitation of benzo-chloro-hexadienone induce the dramatically changes of the SPR signal.Through competition with biotin-RNA for binding site on SPR chip, miRNA attach on the chip.The SPR signal cut down because of the concentration of SA-HRP on the chip decrease. Accroding to this competition principle,we introduce a novel way to detect microRNA.