气助磁分离技术用于蛋白质规模化连续分离的应用研究
发布时间：2018-08-08 17:35
【摘要】：磁分离技术已经有很多年的发展历史,因分离方法简便快捷、目标性强而广泛应用在生物分离、废水处理等领域中。但是由于该技术难以实现放大和连续化,目前在工业中还尚未推广应用。本文通过将浮选技术耦合到磁分离技术中,发展了新型气助磁分离技术。以3.3μm的磁性聚甲基丙烯酸缩水甘油酯微球(MPNs)用于牛血清白蛋白BSA的分离作为模拟体系,开展了间歇、连续和全流程连续气助磁分离过程的研究,研究内容主要包括下列几个方面:1)采用分散聚合法制备磁性聚甲基丙烯酸缩水甘油酯微球,在微球表面修饰亲和配基,以实现目标蛋白的亲和吸附分离。考察了蛋白质的初始浓度、溶液pH、离子强度等因素对蛋白吸附性能的影响,获得蛋白质的最佳吸附条件。2)在磁性载体的最佳吸附条件下,考察并优化了萃取、洗涤、反萃个单元过程的主要工艺参数。考察了溶液的pH值、气体流速、装载体积等因素对分离效率的影响,对于含磁颗粒浓度为0.72 mg/mL的料液,在pH值为7.2的条件下,装载体积为250 mL,气体流速为60 mL/min,其分离速率为410 mL/min,回收率达98%。其结果表明:间歇条件下,气速能大大加速磁分离过程且能实现磁颗粒的远距离捕获。与单纯的磁分离相比,其对磁场的依赖性大为降低,并且分离过程易于放大。同时也证明该分离过程不会导致蛋白质从磁颗粒表面脱落,也不会引起蛋白质构象的变化。3)在此基础上,进一步开展了连续气助磁分离过程的研究,当气速为250mL/min时,其处理量达24 L/h,95%以上的磁颗粒从上部磁辊流出,底部流出液中磁颗粒的浓度仅为0.0389 mg/m L,整个过程质量基本守恒。该过程顺利实现了生物分离过程中磁性微球的规模化连续分离。4)基于上述各单元过程的研究,将全流程规模化连续气助磁分离中试装置用于蛋白质萃取、洗涤和反萃过程的连续化分离,证明了该过程的可行性和高效性。
[Abstract]:Magnetic separation technology has been developed for many years. It has been widely used in biological separation, wastewater treatment and other fields because of its simple and fast separation method and strong target. However, because the technology is difficult to realize amplification and continuity, it has not yet been popularized and applied in industry. In this paper, a new gas-assisted magnetic separation technology is developed by coupling flotation technology with magnetic separation technology. Using 3.3 渭 m magnetic polyglycidyl methacrylate microspheres (MPNs) as a simulation system for the separation of bovine serum albumin (BSA) BSA, the intermittent, continuous and whole-process continuous gas-assisted magnetic separation processes were studied. The main contents of this study are as follows: 1) Magnetic polyglycidyl methacrylate microspheres were prepared by dispersion polymerization. The affinity ligands were modified on the surface of the microspheres to achieve the affinity adsorption separation of the target proteins. The effects of the initial concentration of protein, pH of solution and ionic strength on the adsorption properties of protein were investigated. The optimum adsorption conditions of protein were obtained. 2) under the optimum adsorption conditions of magnetic carrier, the extraction and washing of protein were investigated and optimized. The main process parameters of the stripping unit process. The effects of pH value of solution, gas flow rate and loading volume on the separation efficiency were investigated. When the concentration of magnetic particles was 0. 72 mg/mL, the pH value was 7. 2. The loading volume is 250 mL, the gas flow rate is 60 mL / min, the separation rate is 410 mL / min, and the recovery is 98%. The results show that gas velocity can greatly accelerate the magnetic separation process and realize the long distance capture of magnetic particles under intermittent conditions. Compared with simple magnetic separation, its dependence on magnetic field is greatly reduced, and the separation process is easy to amplify. It is also proved that the separation process will not cause protein to fall off the surface of magnetic particles or cause the conformation of protein to change. 3) on this basis, the continuous gas-assisted magnetic separation process has been further studied. When the gas velocity is 250mL/min, More than 95% of the magnetic particles were disposed of from the upper magnetic roller, and the concentration of the magnetic particles in the bottom effluents was only 0.0389 mg/m / L. the quality of the whole process was basically conserved. This process successfully realized the large-scale continuous separation of magnetic microspheres in biological separation. Based on the research of each unit process mentioned above, the whole process of large-scale continuous gas-assisted magnetic separation (GASM) was applied to the extraction of protein. The continuous separation of washing and stripping process proves the feasibility and efficiency of the process.
【学位授予单位】：湘潭大学
【学位级别】：硕士
【学位授予年份】：2015
【分类号】：TQ028;O629.73
本文编号：2172517
[Abstract]:Magnetic separation technology has been developed for many years. It has been widely used in biological separation, wastewater treatment and other fields because of its simple and fast separation method and strong target. However, because the technology is difficult to realize amplification and continuity, it has not yet been popularized and applied in industry. In this paper, a new gas-assisted magnetic separation technology is developed by coupling flotation technology with magnetic separation technology. Using 3.3 渭 m magnetic polyglycidyl methacrylate microspheres (MPNs) as a simulation system for the separation of bovine serum albumin (BSA) BSA, the intermittent, continuous and whole-process continuous gas-assisted magnetic separation processes were studied. The main contents of this study are as follows: 1) Magnetic polyglycidyl methacrylate microspheres were prepared by dispersion polymerization. The affinity ligands were modified on the surface of the microspheres to achieve the affinity adsorption separation of the target proteins. The effects of the initial concentration of protein, pH of solution and ionic strength on the adsorption properties of protein were investigated. The optimum adsorption conditions of protein were obtained. 2) under the optimum adsorption conditions of magnetic carrier, the extraction and washing of protein were investigated and optimized. The main process parameters of the stripping unit process. The effects of pH value of solution, gas flow rate and loading volume on the separation efficiency were investigated. When the concentration of magnetic particles was 0. 72 mg/mL, the pH value was 7. 2. The loading volume is 250 mL, the gas flow rate is 60 mL / min, the separation rate is 410 mL / min, and the recovery is 98%. The results show that gas velocity can greatly accelerate the magnetic separation process and realize the long distance capture of magnetic particles under intermittent conditions. Compared with simple magnetic separation, its dependence on magnetic field is greatly reduced, and the separation process is easy to amplify. It is also proved that the separation process will not cause protein to fall off the surface of magnetic particles or cause the conformation of protein to change. 3) on this basis, the continuous gas-assisted magnetic separation process has been further studied. When the gas velocity is 250mL/min, More than 95% of the magnetic particles were disposed of from the upper magnetic roller, and the concentration of the magnetic particles in the bottom effluents was only 0.0389 mg/m / L. the quality of the whole process was basically conserved. This process successfully realized the large-scale continuous separation of magnetic microspheres in biological separation. Based on the research of each unit process mentioned above, the whole process of large-scale continuous gas-assisted magnetic separation (GASM) was applied to the extraction of protein. The continuous separation of washing and stripping process proves the feasibility and efficiency of the process.
【学位授予单位】：湘潭大学
【学位级别】：硕士
【学位授予年份】：2015
【分类号】：TQ028;O629.73
【参考文献】
相关期刊论文 前1条
1 陈代雄;复杂多金属硫化矿中铜铅浮选分离工艺研究[J];有色金属(选矿部分);1997年02期
，本文编号：2172517
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