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Method for purifying mouse herve growth factor for scale-production

A technology of nerve growth factor and purification method, which is applied in the field of preparation of mouse nerve growth factor, can solve the problems of affecting production efficiency, increasing protein pollution, long production cycle, etc., and achieves the goal of improving production efficiency, fast flow rate and high load Effect

Active Publication Date: 2008-04-30
ANHUI BIOCHEM BIO PHARMA +1
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Problems solved by technology

[0005] Carboxymethylcellulose cationic chromatography medium (CM-52) was used in this preparation process. Due to the slow flow rate of this medium during use, the height of the column bed will change with the buffer concentration and pH, and it cannot withstand 0.1 NaOH above M is cleaned in place, and the regeneration effect is poor. If it is used in large-scale production, it will affect the production efficiency.
In addition, in the prior art, the mouse submandibular gland homogenate and I-column flow-through both need to be dialyzed for 24 hours, which makes the production cycle too long and increases the possibility of protein contamination

Method used

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  • Method for purifying mouse herve growth factor for scale-production
  • Method for purifying mouse herve growth factor for scale-production
  • Method for purifying mouse herve growth factor for scale-production

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Embodiment

[0023] A method for purifying mouse nerve growth factor suitable for large-scale production, the specific operation method is as follows:

[0024] 1. Submandibular Gland Treatment

[0025] Take out the frozen submandibular gland of male mice, after thawing, add 5 times pre-cooled high-pressure sterilized water, and put it into a homogenizer for homogenization. After homogenization, the homogenate was allowed to stand at 4°C for 30 minutes, centrifuged at 12,000 rpm for 30 minutes, and the supernatant was collected. The supernatant was ultrafiltered to pH 6.8 with a hollow fiber column with a molecular weight cut-off of 10K, and placed in a 20 mmol / L PB solution.

[0026] 2. Sepharose XL (SP XL) (Sepharose SP XL column) chromatography

[0027] The ultrafiltered supernatant was loaded onto a Sepharose XL cation-exchange column (XK 26 / 50, Amersham Biosciences) at pH 6.8, 20mmol / L PB solution equilibrated, flow rate 10ml / min, and the flow-through was collected. See figure 1 . ...

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Abstract

The invention relates to the manufacture method for mouse nerve growth factor. The feature is that ultra-filtrating the supernatant of homogenated and centrifuged mouse submaxillary gland into pH value 6.8, 20mmol / L phosphate buffer solution through hollow fiber column. The Sepharose FF column would take chromatography to collect target albumen, and the Superdex 75prepgrade column collecting the target albumen to gain the mouse nerve growth factor raw liquor. The invention shortens the producing period and improves production efficiency.

Description

technical field [0001] The invention relates to the technical field of biomedicine extraction, in particular to a preparation method of mouse nerve growth factor (NerveGrowth Factor, NGF). Background technique [0002] Nerve Growth Factor (NGF) is the only biologically active molecule found so far that not only acts as a trophic factor on normal nerve cells in the central and peripheral nerves, but also regulates the repair function of damaged nerves. Nerve growth factor has significant clinical application value. The current indications mainly include various peripheral neuropathies, optic nerve injury, spinal injury, craniocerebral injury, damage caused by central nervous system hypoxia and ischemia, and Alzheimer's disease. : NGF can be extracted from the submandibular gland of male mice, human fetal brain, human placenta and snake venom, and can also be prepared by genetic engineering. Among them, NGF expressed by genetic engineering is a development trend, but because ...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C07K14/48C07K1/34
Inventor 陈薇于颖群付玲王传生徐晓红黄小枫
Owner ANHUI BIOCHEM BIO PHARMA
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