Purification method for human chorionic gonadotropin
A chorionic gonadotropin and purification method technology, applied in the field of biopharmaceuticals, can solve problems such as unsatisfactory effects, and achieve the effects of simple and easy operation of the purification process, improved yield, and optimized purification process.
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Embodiment 1
[0033] Embodiment 1: a kind of purification method of human chorionic gonadotropin, such as figure 1 , the specific operation steps are as follows:
[0034] S1, take 1 kg of crude human chorionic gonadotropin with a potency of 200 IU / mg, dissolve it in 50 L of water, add 0.1 kg of polyethylene glycol, stir and centrifuge at 3800 rpm, 8°C for 25 min, use 0.65 μm, 2.5 inch filter cartridge, and adjust the pH=7 of the filtrate with acetic acid;
[0035] S2, the filtrate obtained in step S1 is purified by 5L HS gel hydrophobic column chromatography, fully washed with 45L, 0.5mol / L NaCl washing solution to remove foreign proteins, and washed with 8L, 0.2mol / L NaCl Eluting with liquid, collecting about 8L of active ingredients;
[0036] S3, perform monoclonal antibody affinity chromatography on the active ingredient obtained in step S2, wash fully with 3L, 0.5mol / L NaCl washing solution to remove impurities, and use 8L, 0.1mol / L HCl eluent to elute , collect about 2L of active in...
Embodiment 2
[0038] Embodiment 2: A kind of purification method of human chorionic gonadotropin, the specific operation steps are as follows:
[0039] S1, take 1 kg of crude human chorionic gonadotropin with a potency of 200 IU / mg and dissolve it in 50 L of water, add 0.1 kg of polyethylene glycol, stir and centrifuge at 4000 rpm and 10°C for 20 min, use 0.65 μm, 2.5 inch filter cartridge, and adjust the pH=7 of the filtrate with acetic acid;
[0040] S2, the filtrate obtained in step S1 is purified by 5L HS gel hydrophobic column chromatography, fully washed with 50L, 0.5mol / L NaCl washing solution to remove foreign proteins, and washed with 10L, 0.2mol / L NaCl Eluting with liquid, collecting about 10L of active ingredients;
[0041] S3, perform monoclonal antibody affinity chromatography on the active ingredient obtained in step S2, wash fully with 5 L, 0.5 mol / L NaCl washing solution to remove impurities, and use 10 L, 0.1 mol / L HCl eluent to elute , collect about 2L of active ingredie...
Embodiment 3
[0043] Embodiment 3: A kind of purification method of human chorionic gonadotropin, the specific operation steps are as follows:
[0044] S1, take 1kg of crude human chorionic gonadotropin with a potency of 200IU / mg and dissolve it in 50L of water, add 0.1kg of polyethylene glycol, stir and centrifuge at 4200rpm, 15°C for 18min, use 0.65μm, 2.5 inch filter cartridge, and adjust the pH=7 of the filtrate with acetic acid;
[0045]S2, the filtrate obtained in step S1 is purified by 5L HS gel hydrophobic column chromatography, fully washed with 55L, 0.5mol / L NaCl washing solution to remove foreign proteins, and washed with 12L, 0.2mol / L NaCl The eluent was eluted, and about 12L of active ingredients were collected;
[0046] S3, perform monoclonal antibody affinity chromatography on the active ingredient obtained in step S2, wash fully with 8L, 0.5mol / L NaCl washing solution to remove impurities, and use 12L, 0.1mol / L HCl eluent to elute , collect about 3L of active ingredients, ...
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