The reason why one can be extracted from different types of soil is that different situations are different in their many physical, chemical and biological properties. These are different due to the sequence and clues of the clues from different backgrounds. Connected to the connecting residues on the main chain, positively charged, negatively charged, attracted or non-attractive, attracted or formed, and the connection can be folded into a very good structure (α angle, β sheet and Various rotations, tertiary structures and quaternary structures form unique sizes, shapes and distributions of bases on the surface. Using the difference in properties between the separated parts and other surfaces, methods can be used to observe the confrontation of different properties, namely Design a series of reasonable subdivision steps.
1. Molecular size
There’s a variety of different molecules that have some space in terms of the size of the molecules, and you can use some lens methods to separate the details.
1.1. Rules and ultrafiltration
Some are commonly used in refining to remove salts (desalting and replacement fluids), organic solvents, low molecular concentrations, etc.Ultrafiltration is commonly used to reduce and displace solutions.
1.2. Separation and replacement buffer
The enzyme is enriched in a certain organelle, and then separated into a subcellular component by different plasma. The enzyme is enriched 10-20, and then the specific enzyme is modified.Velocity zone method, if the centrifugation time is too long, all substances will precipitate, so it is necessary to choose the best separation time, can get quite pure subcellular components for further refining, avoid the difference from the heart of the size of the components together with the problem of minutes, can only be used for a small amount of preparation.
1.3. Filter (GF)
Different drugs can be used for desalting, buffering and the removal of drug diversity.The heat source.
The shape of the particles will be affected by the centrifugal movement, the filtration of the particles when passing through the membrane, the movement of the filter, the movement of the filter, or the movement of the small holes on the electrode. Effective radius (Stoke radius), the friction force encountered when passing through the environment is small, the speed is fast and fast, and the color is larger than other shapes; on the contrary, at the peak volume exclusion, the spherical feature of the Stoke radius is easier to penetrate Filter filters are easier to seep out of the surface and have smaller details than other shapes.
There are many ways to affect the solubility of various substances, including: the pH value, ionic strength, dielectric constant and temperature of the solution, but under different specific substance conditions, the main ones have different solubility.
3.1. pH control and isoelectric point precipitation
Proteins are generally more difficult to dissolve at their isoelectric point.
3.2. Organic product separation method
The solubility of microorganisms in different solvents varies greatly, ranging from basically insoluble (<10ug/ml) to extremely soluble (>300mg/ml). The concentration of the organic solvent introduced into the microorganisms is different, so the concentration of the organic solvent can be controlled to polymerize. Dissolving non-ionic polymers such as poly can also cause spots.
In most cases, it is relatively stable at low temperatures, so it is generally operated at 0°C or simulated temperature.
With the net edge of the negative impact at the bottom of the total, and the neutral under the net negative event band then the event occurs.
At isoelectric focusing depth, pI has a difference of 0.02pH.To 100,000 egg whites.
4.2 Ion Exchange Layer (IEX)
In particular mixed solution salt ion strength, pH value and (Yin and Yang) ion exchange device, different phenomena to different ion exchange failure to prevent different ability, different properties or observed separation.
Siphoning can be done either by keeping the adsorbent composition constant or by changing the salinity or pH of the agent, which can then be converted to expression and dripping for the dripping effect.Exquisite, exquisite, especially the ion exchanger with small exchange capacity and sensitivity to salt concentration, multi-purpose rainbow color.Can be separated from the ion exchange column.
5.1 Layer by Layer Analysis (HIC)
Most marginal marginal residues are hidden in the interior, but there are also some surfaces.The surface aspect of the surface affects the edges and spatial propagation of residues and determines whether the structure has the characteristics of the microbe that binds to the edges and uses them for refinement and refinement. It is a universal tool for protein separation and purification. Therefore, it is especially suitable for the mother solution after concentrated sulfate solution precipitation separation and the solution containing the product after precipitation dissolved with salt directly into the sample column, of course, also suitable for 7mol/L hydrochloric acid or 8mol/L large intestine therapeutic antigen at the same time leaching solution directly into the sample column, in the separation is also renatured.
6. Screenshot of Affinity Layer (AC)
Combines the characteristics of high efficiency and fast separation speed. The base ligand can be the substrate, auxiliary, auxiliary feature, and feature of the enzyme. After the removal, the strength and pH value of the buffer can be changed. Observed, the concentration of the same ligand solution or particle distribution of the ligand solution can also be increased. The different ligands can be divided into:
6.1. Metal electrode combination
The transition metal ions Cu2+, Zn2+ and Ni2+ are bonded to the stationary phase in the form of imine complex ions. Because these metal ions form valence bonds with tryptophan, histidine and semi-possibility, the image is stabilized. The performance is controlled by a single histidine and semi-material dissociation constant, which can also be affected by the pH value and temperature of the mobile phase, and the control conditions can cause different breakages.
6.2 small ligand affinity
Equipped with arginine, benzosomes, calmodulin, gelatin, heparin and lysine, etc.
6.3. Oppose affinity
That is, immunoaffinity layer binding, where the ligand has A set of A and A set of native protein G, but protein A is more specific than protein G, and protein G can bind to many different types of IgG.
6.4. Picture affinity
In addition to the size of ligands and enzymes, the effect of a layer of contrast is also related to the type of drug solution, ionic strength, pH value and the details to be separated..Under certain conditions, the immobilized can stimulate the action of ac exchangers to avoid this phenomenon, the best ionic strength is less than 0.1 and pH value is greater than 7 when the operation.
6.5. Exogenous lectin affinity medium
The ligand has the properties of concanavalin, the binding energy of exogenously lectin of bean and lectin of soybean and several kinds of carbohydrate residues, and is suitable for refining polysaccharides and glycoproteins.
7.1. thermal stability
All heated images will unfold or precipitate when heated to 95°C, which can easily separate a kind of particles that remain granular after heating from other cellular materials.
7.2. simulation solution stability
Treat the supernatant with cancer, chaperone protein, and anti-deteriorating resistance together.