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Preparation method of porcine insulin

A technology for insulin and protein precipitation, which is applied in the preparation methods of peptides, insulin, chemical instruments and methods, etc., can solve the problems of limited organic solvents in the production process, low purity of pig insulin, etc., and achieves improved commercial purity and optimized preparation process. , the effect of reducing production costs

Pending Publication Date: 2021-06-22
江苏万邦医药科技有限公司
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

The purity of commercial porcine insulin is relatively low, and the production process involves limited organic solvents

Method used

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  • Preparation method of porcine insulin
  • Preparation method of porcine insulin

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0044] reversed phase chromatography

[0045] The loading sample is the precipitated protein precipitated by adding zinc after ion exchange chromatography. The precipitated protein was redissolved according to the ratio of protein amount to solution volume 1:10 (g / ml), pH=3.5.

[0046] Reverse-phase exchange chromatography was performed on a Relianx high-pressure chromatography system using UniPS 10-300 packing material. Described buffer solution A is the buffer solution that mass fraction is 10% ethanol and 100mmol / L sodium chloride formation; Described buffer solution B is the buffer solution that mass fraction is 50% ethanol and 100mmol / L sodium chloride formation, All solutions were adjusted to pH 3.0 with hydrochloric acid. The column volume of the chromatography column is 20ml. The buffer gradient is 0-60% B 1CV, 60-80% B 15CV, and the linear flow rate is 152cm / h. The detection wavelength is 280nm.

[0047] The specific steps are:

[0048] (1) Buffer A liquid equil...

Embodiment 2

[0054] The loading sample is the same as the sample in Example 1.

[0055] Reverse-phase exchange chromatography was performed on a Relianx high-pressure chromatography system using UniPS 10-300 packing material. Described buffer A is the buffer that massfraction is 10% ethanol and 100mmol / L sodium chloride; Described buffer B is the buffer that massfraction is 50% ethanol and 100mmol / L sodium chloride, All solutions were adjusted to pH 3.5 with hydrochloric acid. The column volume of the chromatography column is 20ml. The buffer gradient is 0-60% B 1CV, 60-80% B 15CV, and the linear flow rate is 152cm / h. The detection wavelength is 280nm.

[0056] The specific steps are:

[0057] (1) Buffer A liquid equilibrium packing 2CV (CV is the abbreviation of Column Volume, which is the column bed volume of the purification column)

[0058] (2) Take about 200mg of sample and load it, and continue to equilibrate 2CV with buffer A solution after loading

[0059] (3) Buffer A and B ...

Embodiment 3

[0063] The loading sample is the same as the sample in Example 1.

[0064] Reverse-phase exchange chromatography was performed on a Relianx high-pressure chromatography system using UniPS 10-300 packing material. Described buffer A is the buffer that massfraction is 10% ethanol and 100mmol / L sodium chloride; Described buffer B is the buffer that massfraction is 50% ethanol and 100mmol / L sodium chloride, All solutions were adjusted to pH 4.0 with hydrochloric acid. The column volume of the chromatography column is 20ml. The buffer gradient is 0-60% B 1CV, 60-80% B 15CV, and the linear flow rate is 152cm / h. The detection wavelength is 280nm.

[0065] The specific steps are:

[0066] (1) Buffer A liquid equilibrium packing 2CV (CV is the abbreviation of Column Volume, which is the column bed volume of the purification column)

[0067] (2) Take about 200mg of sample and load it, and continue to equilibrate 2CV with buffer A solution after loading

[0068] (3) Buffer A and B ...

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Abstract

The invention relates to a preparation method of porcine insulin, which comprises the following steps: performing ion exchange and reversed phase chromatography purification on porcine insulin extracted from porcine pancreas to prepare the porcine insulin with the purity of more than 99%, and performing crystallization, washing and drying to obtain a porcine insulin finished product. The purity of the porcine insulin purified by ion exchange chromatography and the yield of the porcine insulin purified by reverse phase chromatography can reach 99% and 92%, and the purity of the finished product obtained after crystallization, washing and drying reaches 99%. The purity of the porcine insulin is further improved, the content of related impurity proteins is reduced, meanwhile, the related organic solvent is only industrial ethanol which is low in price and easy to obtain, the production cost is reduced, and the product quality is improved.

Description

technical field [0001] The invention relates to the fields of drug synthesis and chemical industry, in particular to a preparation method of porcine insulin. Background technique [0002] Diabetes is a common metabolic endocrine disease, which has become the third chronic non-communicable disease that threatens human health after cardiovascular and cerebrovascular diseases and malignant tumors. Diabetes is often accompanied by a variety of complications, which can easily cause multiple organ damage, functional impairment and failure, and is very harmful. At present, China has become the country with the largest number of diabetic patients in the world. [0003] Insulin is one of the most effective drugs for diabetes treatment, and porcine insulin is extracted from pig pancreas, which is convenient and easy to obtain, and at the same time, the price is cheaper than other insulins, which can meet the needs of some impoverished areas. The purity of commercially available porc...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C07K14/62C07K1/36C07K1/30C07K1/20C07K1/18
CPCC07K14/62
Inventor 文良柱姜皓支艳艳赵珊珊
Owner 江苏万邦医药科技有限公司
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