Protein separation and purification in genetic engineering workshop
Protein separation and purification in genetic engineering workshop
Protein separation and purification is the core technology in the biological industry and an important link in the pharmaceutical industry of genetic engineering workshops; the basic principles of separation and purification of recombinant proteins are capture, crude purification, and purification. The general process of separation and purification is : First of all, the biological material is pre...
Text label: genetic engineering workshop, protein separation and purification, genetic engineering pharmaceutical cleanroom construction
Protein isolation and purification in genetic engineering workshop
Protein separation and purification is the core technology in the biological industry and an important link in the pharmaceutical industry of genetic engineering workshops; the basic principles of separation and purification of recombinant proteins are capture, crude purification, and purification. The general process of separation and purification is : First pre-process the biological material to separate the recombinant protein from the cells, and then select an appropriate separation and purification method according to the properties of the protein to be purified, such as the isoelectric point of the protein, solubility in different salt solutions, etc. Precipitation separation method can be used for properties; membrane separation technology can be used according to protein molecular weight; ion exchange chromatography, molecular sieve chromatography, hydrophobic layer can be selected according to charge difference, molecular size, hydrophobicity and specificity of affinity with other biomolecules Chromatography and Chromatography.
Pretreatment of protein samples, the pretreatment process generally involves the separation or enrichment of cells or bacteria, and the methods of centrifugation and filtration are used, which requires the state of the fermentation broth and culture broth to improve the solid-liquid separation. Speed; the pretreatment of the fermentation broth or culture broth of genetically engineered strains and cells is to reduce the viscosity of the solution by adjusting PH, heating and other treatments under the premise of ensuring the stability of the biological activity of the protein, changing the properties of the solution, and making the state more conducive to Follow-up action. Some of the products expressed by genetically engineered strains and cells can be secreted outside the cells, but most of them exist in the cells. Only after the cells are broken can the products in the cells be released to the liquid phase to the greatest extent for further extraction. The methods of cell fragmentation include mechanical fragmentation method and non-mechanical fragmentation method; non-mechanical fragmentation method mainly uses physical factors and biochemical substances to break cells, mainly including freeze-thaw method, chemical lysis method, enzymatic hydrolysis method, etc.; mechanical fragmentation method It mainly uses the mechanical force of the liquid and solid phases to break the cells. The homogenate after cell fragmentation contains a large amount of cell debris, which must be separated before further extraction and separation. At present, methods such as filtration, centrifugation, and aqueous phase extraction are mainly used. Zhongjing Global Purification can provide consulting, planning, design, construction, installation, transformation and other supporting services for clean workshops and genetic engineering clean workshops.
Membrane separation technology is a fluid separation operation technology represented by polymer separation membranes. Because membrane separation technology has many advantages that other separation technologies do not have: concentration does not require phase change, separation coefficient is large, separation selectivity is strong, and it does not require Evaporator or refrigeration equipment, low investment cost and low energy consumption; membrane separation technology has many advantages and importance, and is widely used in biological and food industry, environmental engineering, pharmaceutical industry and many other aspects. Membrane separation technology uses a semi-permeable membrane as a selective barrier layer. There is a certain amount of energy difference on both sides of the membrane as a driving force, allowing some components to pass through while retaining other components in the mixture, and the migration of each component head through the membrane. It is a technology that achieves the purpose of separation with different rates; the membrane is a material with selective separation function, and the selective separation of the membrane is used to achieve the separation, purification and concentration of different components of the feed liquid. It is different from traditional filtration in that the membrane can Separation is carried out in the molecular range, and this process is a physical process, which does not require phase change and the addition of additives. The pore size of the membrane is generally micron. Ultrafiltration membranes, nanofiltration membranes and reverse osmosis membranes; according to different materials, they can be divided into inorganic membranes and organic membranes. Inorganic membranes are mainly ceramic membranes and metal membranes, which have low filtration accuracy and low selectivity.
Ultrafiltration is a membrane process between microfiltration and nanofiltration. The membrane pore size is between 0.05-1000μm. Ultrafiltration is a membrane separation technology that can purify, separate and concentrate solutions. The pressure difference on both sides of the membrane is the driving force, and the ultrafiltration membrane is used as the filter medium. Under a certain pressure, when water flows through the surface of the membrane, only water and small molecular substances smaller than the membrane pore size are allowed to pass through to achieve the purification of the solution, separation and concentration purposes. Microfiltration belongs to precision filtration. The basic principle is the sieve separation process. The application scope of microfiltration membrane is mainly to intercept particles, bacteria and other pollutants from the gas phase and liquid phase, so as to achieve the purpose of purification, separation and concentration. Nanofiltration is a membrane separation technology between ultrafiltration and reverse osmosis. Its molecular weight cutoff is in the range of 80-1000Da, and the pore size is several nanometers. Based on the superiority of nanofiltration separation technology, it is widely used in biochemical, pharmaceutical, Food industry and many other fields have broad application prospects.
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