Purification method of growth hormone
A purification method and technology of growth hormone, which is applied in the field of purification of growth hormone, can solve problems such as complex operation, high cost, and long operation time, and achieve the effects of simplified operation steps, low cost, and improved yield
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Embodiment 1
[0062] After the human growth hormone recombinant Escherichia coli was fermented, the sediment (i.e., the sludge) was collected by centrifugation, placed in a -20°C refrigerator for 24 hours, and then thawed at 20-25°C for 12 hours. The extract was added according to the ratio of bacteria slime:extract = 1:14 (W:W), and the extract was an aqueous solution containing 100 mM glycine and 1 mM edetate disodium at pH 7.0. Microfiltration is carried out through a 1000kDa hollow fiber membrane, and then the filtrate obtained by the microfiltration is subjected to ultrafiltration and concentration through a 10kDa ultrafiltration membrane to obtain a retentate. Add sodium chloride to the retentate at a concentration of 2.0 mol / L, adjust the pH to 6.0-6.5, and filter through a 0.45 μm filter membrane. Rinse the hydrophobic chromatography column filled with PhenylHP for 8 to 10 CVs with an equilibrium solution (aqueous solution containing 10mM sodium dihydrogen phosphate and 2.0mol / L sod...
Embodiment 2
[0064] After the human growth hormone recombinant Escherichia coli was fermented, the precipitate (i.e., the sludge) was collected by centrifugation, placed in a -50°C refrigerator for 36 hours, and then thawed at 20-25°C for 14 hours. Add the extract solution according to the ratio of bacteria sludge: extract solution = 1:10 (W:W), and the extract solution is 5 mmol / L sodium dihydrogen phosphate aqueous solution. Microfiltration is carried out through a 1000kDa hollow fiber membrane, and then the filtrate obtained by the microfiltration is subjected to ultrafiltration and concentration through a 10kDa ultrafiltration membrane to obtain a retentate. Add sodium chloride to the retentate at a concentration of 2.5 mol / L, adjust the pH to 6.0-6.5, and filter with a 0.45 μm membrane. Rinse the hydrophobic chromatography column with Phenyl HP filler for 8 to 10 CVs with equilibrium solution (aqueous solution containing 10mM disodium hydrogen phosphate and 2.0mol / L sodium chloride) a...
Embodiment 3
[0066]After the human growth hormone recombinant Escherichia coli was fermented, the sediment (i.e., the sludge) was collected by centrifugation, placed in a -80°C refrigerator for 30 hours, and then thawed at 20-25°C for 16 hours. According to the sludge: extraction solution = 1:20 (W: W), add the extraction solution, the extraction solution is 50mmol / L sodium dihydrogen phosphate aqueous solution. Microfiltration is carried out through a 500kDa hollow fiber membrane, and then the filtrate obtained by the microfiltration is subjected to ultrafiltration and concentration through a 10kDa ultrafiltration membrane to obtain a retentate. Add sodium chloride to the retentate at a concentration of 3.0 mol / L, adjust the pH to 6.0-6.5, and filter through a 0.22 μm filter membrane. The hydrophobic chromatography column filled with Phenyl HP was first washed with equilibrium solution (aqueous solution containing 10mM disodium hydrogen phosphate and 3.0mol / L sodium chloride) for 8 to 10 ...
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