Research On A New Preparative Vertical Flow Electrophoresis System With Supporting Medium

Electrophoresis technology is an important method for separation and purification.Because of its higher resolution,recovery rate,and wider exploitability,it has played an important role in the separation and detection of many types of substances(biological and non-biological).Since the development of electrophoresis technology,most of them are miniature or discontinuous,and there are few studies on electrophoresis devices for the purpose of separation and preparation.Therefore,this paper attempts to propose a preparative vertical flow electrophoresis that can perform continuous separation.By filling a certain supporting medium,it reduces the convection diffusion that is easy to occur in the vertical flow process.That is,a preparative vertical flow electrophoresis device with a supporting medium.Through the operation design,a preparative vertical flow electrophoresis system with a supporting medium is formed,Through the investigation of the operation stability and separation preparation of the system,the usable value of the system is proved.In the system design,it mainly includes seven parts: preparative vertical flow electrophoresis device,sample sampling system,background buffer sampling system,electrode buffer circulation system,cooling circulation system,sample collection system and electrophoresis power supply;in the design of the device The main structure consists of the separation chamber,ion exchange membrane,electrode chamber and cooling circulation water inlet chamber.In the investigation of the stability of the system,the bromophenol blue was used as the sample under no voltage,and the movement trajectory in the separation chamber was observed.The movement was linear,and the inlet and outlet basically corresponded,indicating that there was no leakage and large deviation of the system.Under applied voltage,the study shows that the voltage-current curve within 175 V has a good linear effect.In the investigation of the upper voltage limit of the device,the upper limit voltage is 175 V.After investigating the stability of the system,through the experimental design,the system was used in the form of zone electrophoresis and continuous sampling to achieve the separation of mixed proteins(Bovine Serum Albumin and Cytochrome C),demonstrating the device's Practical value.In order to obtain better separation effect and resolution,under 100 V voltage,optimized the experimental conditions for separation of mixed proteins: The optimal background buffer flow rate is 0.183 mL/min,the optimal background buffer concentration is 40 mM,and the optimal sample flow rate is 0.04 mL/min.Preliminary research shows that the self-designed preparative vertical flow electrophoresis system with supporting medium can achieve simple mixed protein separation,and it also has certain application potential in the separation of complex proteins and even other charged substances;For example,excess sludge as a misplaced resource,The protein in the hydrolysate obtained by alkaline digestion of the remaining sludge occupies a large proportion,and the nucleic acid polysaccharide also occupies a large proportion.These biological macromolecules generally have two properties of acid and alkali,and have different mass-to-charge ratios under certain conditions.The separation and recovery of these substances can be achieved through the electrophoresis process,which has great significance for the reuse of residual sludge.With the foundation of electrophoresis,the device has a large space for development.