Purification of rHSA

A technology of exchange chromatography and cation exchange, which is applied in the field of protein purification, can solve the problems of high levels of other impurities, and achieve the effects of short experimental period, good application prospects and high product purity

Active Publication Date: 2006-11-01
NCPC NEW DRUG RES & DEV
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  • Abstract
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Problems solved by technology

Compared with other methods, the A350 / 280 ratio of the chromogenic substance level of the recombinant HSA purified by this method can be reduced to 0.02-0.04, but the level of other impurities is relatively high, and further optimization is needed

Method used

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  • Purification of rHSA
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  • Purification of rHSA

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Experimental program
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Embodiment Construction

[0027] The purification process of rHSA of the present invention is as follows:

[0028] In order to reduce the production of pyrogens, the rHSA fermentation broth was filtered with a 0.22um membrane (manufactured by MILLIPORE), and then heated in time to inactivate protease. In order to inhibit the combination of exogenous pigments and target proteins, ensure protein stability, and inhibit the activity of high-temperature-resistant proteases, add decolorization media, stabilizers, and enzyme inhibitors when heating. Decolorization media such as macroporous resin, diatomaceous earth, and activated carbon etc., preferably activated carbon; stabilizers such as palmitic acid, sodium octanoate, etc., enzyme inhibitors such as PMF, EDTA, EGTA, etc., preferably EDTA. In the simultaneous presence of stabilizers, decolorizing media and enzyme inhibitors, adjust the pH of the fermentation supernatant to 6.0-7.0, and then start heating at a temperature of 60-75°C for 10-100 minutes, pre...

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Abstract

Purification of rHSA is carried out by heat treating for fermented supernatant containing rHSA at 60-75degree under existence of stabilizer, decolorant and proteinase inhibitor, adjusting solution pH4.0-5.5, hypersaline cation exchange chromatographying for fermented supernatant containing rHSA, hydrophobic exchange chromatographying and weak anion exchange chromatographying to obtain purified rHSA and borate treating. It has short experimental period and higher purity.

Description

technical field [0001] The invention relates to a protein purification method, in particular to a method for purifying rHSA. Background technique [0002] Human Sorum Albumin (HSA) is the most abundant protein component in plasma, which helps to maintain blood osmotic pressure, and can combine small molecule nutrients and drugs to play a transport role. Clinically used to treat hypoalbuminemia, protein deficiency syndrome, shock and burns after trauma and surgery, liver cirrhosis, nephrotic syndrome, etc. It is also used as an excipient for other medicines, as a coating for pipelines in medical devices, as a medium for in vitro fertilization, etc. [0003] The earliest HSA obtained has been obtained by plasma separation method. However, this method has the disadvantages of insufficient supply of raw materials, periodic shortages, and carrying hepatitis virus and HIV virus, which not only brings serious harm to operators during the production process, but also has potential...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C07K14/76C07K1/16
Inventor 李梅彦贾茜张炜董爱华任力伟沈旭东高健邓建慧
Owner NCPC NEW DRUG RES & DEV
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