The Separation And Detection Of Elastase With Surface Molecular Imprinting

Protein exists in organisms, and takes part in vital physiological activities, such as construction and repairment of tissue, enhancement of organism immunity, adjustment of osmotic pressure, construction of enzyme and hormone, supply of energy. The needs of high purity have been come up with the investigation in and applying of protein, which make the separation and detection of protein vitally important. However, the protein is derived from organisms therefore bonded with their specialities of high vitality and volatility which make separation of protein complicated and costly. Thus, a gentle and efficient technology of protein separation makes great differences. This thesis is aiming on high-efficient separation of and detection on elastase in order to provide some technical reference for industrial manufacture of elastase. Following is the content of the research.1. Molecular imprinting polyvinylidene fluoride(PVDF) membrane was prepared via using prepolymer to coat PVDF membrane. The experiment was optimized about the effect of the amounts of each components. In addition, its extraction performance was studied through homemade membrane module. The surface groups of PVDF membrane were changed due to the molecular imprinting layer, and the recognition sites of elastase were brought in PVDF membrane, which added the extraction capability of membrane silk to elastase. The extraction capacity of molecularly imprinted membrane silk was 1.29 times than that of ordinary membrane silk, and the selectivity of imprinted membrane silk was enhanced greatly. The analysis of SEM and contact angle demonstrated that the preparation of controllable PVDF membrane was successful and the repeating experiment showed that the imprinted PVDF membrane had excellently reused ability.2. The surface molecular imprinting layer was prepared via using adjacent hydroxyl N-(2-hydroxyethyl)-N-propyl acrylamide as a monomer and N,N-methylene-bis-acrylamide as crosslinking agent on the surface of SiO2 microspheres. Separation performance of microspheres for elastase was studied via investigating the adsorb capacity of imprinted microspheres to elastase. In the paper, imprinted microspheres showed certain selectivity and the adsorb capacity of imprinted microspheres to elastase reached 16mg/g or more. And by TGA and SEM, thin molecular imprinting polymer layer was proved on the surface of silicon microspheres.3. A molecules imprinting fluorescent sensor for detecting elastase was prepared by polymerization on surface of silica microspheres with elastase as a template molecule mixing 1-formaldehyde, TEOS and 3-aminopropyl triethoxysilane. We had investigated the effect of pH, time, the amount of imprinted particles and temperature on the fluorescence response. Meanwhile selectivity and relative standard deviation in the strategy was measured. Using silicon as the main material made this strategy simply preparing, economical and practical.