Glycoprotein Detection Based On Fluorescent Speed Sensing

Chip electrophoresis is a novel micro-total analysis technique based on traditional capillary electrophoresis and has enormous application in the analysis of compounds,drug,proteins and even complicated samples.The chip electrophoresis is apt to be commercialized and has high integration level with fast analysis and low cost.Numerous researches have been reported for glycoprotein detection using micro-chip analysis.However,there is no reports concerning moving reaction boundary?MRB?and fluorescent speed sensing for glycoprotein detection.This thesis advanced the chip supramolecule electrophoresis platform,which takes MRB and fluorescent speed sensing as the core theories.The developed method is based on chip electrophoresis and can be utilized for the glycoproteins and even diabetes detection.The principal work of this study are listed as follows:Synthesis and characterization of the novel supramolecule compound fluorescent probe:We synthesized a novel compound with boronic group?3-?dinaphthopyry-lium-4-yl?phenyl-boronic acid,Rp3?via one-pot reaction,and the Rp3was then simply mixed with fluorescein?F1?to from a novel supramolecule compound fluorescent probe F1@Rp3.A series of characterization and study with respect to F1@Rp3?especially the fluorescent respond to glycoprotein?were carried out to prove its feasibility for glycoprotein detection.Fluorescent speed sensing theory:With the purpose to establish a simple,quick and accurate method for glycoprotein analysis and quantification,we proposed the concept of fluorescent speed sensing theory based on the study of multipath moving supramolecular boundary electrophoresis for the first time.Further,the model of fluorescent speed sensing was built in virtue of F1@Rp3.By measuring electrophoretic mobility and using the concept of constituent mobility and content,we stepped further into the theory of fluorescent speed sensing theory.The liner relationship between the migration rate of F1 and protein content under electric field was derived.Validation of fluorescent speed sensing theory:To validate the feasibility and accuracy of our proposed theory,we chose IgG and ovalbumin as the model proteins and the fluorescent speed sensing method which relies on chip supramolecule electrophoresis platform was tested.The results demonstrated that with the increasement of protein content,the released fluorescent molecule F1 migrated faster under electric field.Meanwhile,the protein content and the migration rate of F1 showed high linear relationship and low RSD value.The detectable concentration of IgG and ovalbumin in this fluorescent speed sensing method were 10?g/mL and 25?g/mL,respectively.And the minimum detectable content of these two proteins were 8.48 ng and 21.2 ng.The data acquired illustrates that our fluorescent speed sensing method is feasible and correct,and has good linearity,high accuracy and sensitivity as well.The practical application of fluorescent speed sensing theory in clinical samples:We selected HbA_1c as the practical sample in clinical detection to explore the application prospect of fluorescent speed sensing approach.The results indicate that the HbA_1c content and the migration rate of F1 showed linear correlation,which acts as another evidence of the feasibility and validity of fluorescent speed sensing theory.The proposed theory in this paper,undoubtedly,has potential application prospect in clinical test.