Preparation And Characterization Of Poly (Vinyl Naphthalene) Nanoparticles And Primary Investigation On Micro-column Cell Membrane Chromatography

Objective: To prepare polyvinyl naphthalene(PVN)nanoparticles and carboxylation polyvinyl naphthalene(PVN-COOH)nanoparticles then apply their characterization for latex-enhanced turbidimetric immunoassay(LETIA)of beta 2 microglobin(?2-M).To prepare porous coating of silica gel which is based capillaries were support and apply their for cell membrane chromatography.Methods: PVN nanospheres were prepared by emulsion polymerization.Vinylnaphthalene,sodium dodecyl sulfate and carbonate buffer were employed as polymerizable monomer,emulsifier and aqueous phase,respectively.Use sodium acrylate as carboxylation agent to prepare PVN-COOH nanoparticles.The influences of concentrations of SDS?VN?KPS and sodium acrylate were investigated.The nanoparticles were characterized by SEM,Infrared Spectrum analysis,Micro-particular analysis,etc.In addition,the anti-?2-M antibodies were immobilized onto the PVN and PS nanopariticles by physical absorption.The carboxyl on the surface of the PVN-COOH and PS-COOH microspheres was activated by Sulfo-NHS and EDC,and then coupled with ?2-M to preparate ?2-M-LETIA testing reagent.Finally,the testing results were manipulated and analysised by statistic means.To prepare the chromatographic column of cell membrane chromatography followed the three main steps.First,the pretreatment of the capillary column;Next,porous coating of silica gel was immobilized onto a capillary innerwall.Utilize polyethylene glycol(PEG)and urea as hole making agent,Methyl silicate(TMOS)as the comonomer,glacial acetic acid as the solvent,and the porous coating of silica gel which is based capillaries was fabricated by hydro-thermal reaction ? acid activation and high-temperature drying.Finally immobilizetion of Biological cell membrane(The red blood cells of rabbit are used as the mode).The capillary columns were characterized by High Efficiency Capillary Electrophoresis,Scanning Electron Microscope,Fluorescent labeling method.The verapamil was used as the positive drug.Dynamic binding capacity of mCMC was analysed through the front analysis experiments.Analysis of all the results,then the optimal conditions for preparation of mCMC was established.Results: Via adjusting and controlling of the preparation conditions,the size of the microsphere has been controlled within a certain range.By means of SEM,the nanoparticles have a good degree of sphericility and an uniformitarian size distribution;Carboxyl groups of PVN-COOH naoparitcles is proved by the infrared spectrum.The optimum conditions of coupling ?2-M-antibodies to the PVN?PVN-COOH microspheres have been obtained and the ?2-M-LETIA reagent is prepared.It was compared with PS-?2-M-LETIA reagent,the ?2-M-LETIA reagent has a significant advantage in sensitivity and linear range.The capillaries with porous coating of silica gel were fabricated by the hydrothermal synthesis method.Via changing of the preparation conditions,the thickness ?aperture and pore size distribution of coating are controlled.The stationary phase of mCMC could be visualized by fluorescent labeling of cell membrane.The capacity of cell membrane on different mCMC could be obtained by analyzing of fluorescence intensity.The results about the front analysis experiments of verapamil showed that comparing with the no-coating capillary,DBC of the silica gel-immobilized capillary was improved significantly.What's more,there was significant difference about DBC of different capillaries.The optimum conditions of preparing mCMC capillary have been obtained by analyzing the data of fluorescence intensity and the front analysis experiments of verapamil.Conclusion: The size-controlled and monodispered PVN?PVN-COOH microsphere are fabricated.Development of the method of coupling ?2-M antibodies on nanoparticles and the ?2-M-LETIA reagent is fabricated.After optimization of the coupling conditions,the optimum conditions are obtained.As novel carriers of LETIA reagent,PVN?PVN-COOH nanoparticles have a good application prospect.It was compared with the capillary,the capacity of cell membrane and dynamic binding capacity of positive drug were improved on the silica gel-immobilized capillary.So the silica gel-immobilized capillary was more suitable for mCMC.