Fabrication Of Honeycomb-structured Protein Arrays

This paper combines inverse emulsion system with Breath figure method to prepare protein array structure by one step pattern. The Breath figure, a kind of the self-assembly method, which has been studied for many years, can be simple and effective to implement pattern preparation of array structure, With the organic solvents as the solvent phase to prepare the film liquid, reverse emulsion system introduces a portable water soluble macromolecular components(protein, etc.) as water phase; Then the water as a template evaporating forms the structure of array holes. Protein components can enrich inside the hole of regular arrangement by the process of water phase self-assembly to form a three-dimensional protein pattern array structure eventually. In the early stage of the project, the classic Breath figure method filmforming conditions are optimized to carry on the exploration and the optimum film forming conditions are determined. Then, on the one hand, different ways of formation in order to obtain the stability of inverse emulsion will be explored; on the other hand, many factors will be tried to affect the systematic experiment, such as different water oil ratio, different dosage of emulsifier and the film forming conditions, to seek a stable optimal parameter of inverse emulsion system. The two preparation method of reverse emulsion system, determine the high shear and ultrasonic formation of emulsion, are determined through systematic experiment. In the same conditions, both are relatively more ideal and longer for the preparation of the reverse emulsion stability. Fibronectin and bovine serum albumin protein components are added in the emulsion on the basis of the methods, and filmed on the polyester, glass and silicon. The ordinary optical microscope, fluorescence microscope(FM) and scanning electron microscope(SEM) are studied to obtain much data of surface and cross-section morphology structure. Finally the fluorescein isothiocyanate(FITC) as fluorescent marker to tag protein components in fluorescence microscope and confocal laser scanning microscope(CLSM), which tracks under the characterization of protein components of selective distribution based on preliminary work. The implementation of the project confirmed that the combination of reverse emulsion with Breath figure method is feasible, and the protein pattern array structure is prepared with this method. Biocompatibility polymer materials, such as polylactic acid, polycaprolactone are selected to be membrane substrates bringing in the inverse emulsion system, and combined with biological matrix inverse emulsion(chitosan microspheres) to prepare selectively modified biological film materials with three-dimensional structure of protein array。...