Method for purifying protein
a protein and purification technology, applied in the field of purification methods, can solve problems such as substantial loads, and achieve the effect of reducing loads in the affinity chromatography step
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example 1
[0156](i) Introduction of Graft Chains into Porous Hollow Fiber Membrane
[0157]In an airtight container was placed porous polyethylene hollow fibers having an outer diameter of 3.0 mm, an inner diameter of 2.0 mm and a maximum pore size measured by the bubble point method described in the above (1) of 0.3 μm, and the air in the container was replaced with nitrogen. Then, the hollow fibers were irradiated with 200 kGy of γ-ray to generate radical while cooling the container from the outside thereof with dry ice. The resultant radical-containing porous polyethylene hollow fibers were placed in a glass reaction tube, and the oxygen in the reaction tube was removed by depressurization to 200 Pa or less. A reaction solution containing 3 parts by volume of glycidyl methacrylate (GMA) and 97 parts by volume of methanol, adjusted at 40° C., was injected thereinto in an amount of 20 parts by mass to the hollow fibers and then allowed to stand in a closed state for 12 minutes to subject to gra...
example 2
[0174](iv) Fixation of Anion-Exchange Groups (Quaternary Amino Groups) to Graft Chain
[0175]In the same way as in Example 1, a dried hollow fiber into which graft chains were introduced was swollen by immersion in methanol for 10 minutes or more, and then immersed in purified water for replacement with water. A mixed solution containing 50 parts by volume of purified water and 50 parts by volume of dimethylsulfoxide was provided, and trimethylammonium chloride was added to a concentration of 0.5M to the mixed solution, which was then mixed to provide a uniform reaction solution. The reaction solution was placed in a glass reaction tube in an amount of 20 parts by mass to the hollow fiber after graft reaction, and adjusted to 60° C. The porous hollow fiber into which graft chains were introduced was then inserted thereinto and allowed to stand for 200 minutes to replace the epoxy groups of the graft chains with trimethylamino groups to provide a porous hollow fiber membrane having tri...
example 3
[0179]NaCl was added to 20 mM Tris-HCl (pH 8.0) to a concentration of 0.17 M to prepare a metal salt-containing buffer. In this buffer dissolved were BSA (pI: 5.6) and γ-globulin at a concentration of 1 g / L each to prepare a mixed solution of proteins. This mixed solution of proteins was passed through the anion-exchange membrane module having diethylamino groups prepared in Example 1 at a flow rate of 2 mL / minute, and the filtrate was collected in the form of 5 mL each of fractions. SDS-PAGE was used to analyze proteins in the fractions having passed through the module. The filtrate (10 μL) to be used for the analysis was mixed with an equal amount of a sample treatment solution (Tris SDS sample treatment solution from Daiichi Pure Chemical Co., Ltd.), which was then heat-treated at 100° C. for 5 minutes. The resultant sample was applied to a gel plate for electrophoresis (Multigel II Mini from Daiichi Pure Chemical Co., Ltd.) in an amount of 10 μL per well using a micropipette, wh...
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