Process for rourifying insuline

A purification method and insulin technology, applied in the preparation methods of insulin and peptides, chemical instruments and methods, etc., can solve the problems of increased titer yield, short production cycle, low product yield, etc., and achieve increased titer yield. , yield and purity improvement effect

Inactive Publication Date: 2003-01-15
JIANGSU WANBANG BIOPHARMLS
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Its disadvantages are low product yield, many operation steps, long production cycle, high consumption of auxiliary materials, and about 15% of recycled products are produced in the process at the same time, especially for the production of high-purity insulin. Obviously, its process is unreasonable. Chinese patent Publication No. CN1077203A discloses an invention patent application "a crystallization process of insulin sodium salt".
Compared with the zinc salt crystallization process, this process has fewer procedures and a shorter production cycle, but the disadvantage is that the titer yield is only increased by 3-5%.

Method used

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Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0012] Dissolve 1Kg of insulin salted-out product with 0.1N dilute hydrochloric acid to make a volume of 5 liters, adjust the pH to 8.2 with 10 equivalents of sodium hydroxide, add 0.5 liters of 30% sodium chloride solution, stir evenly, and then use 5N Adjust the pH to 8.2 with sodium hydroxide, stir at room temperature for 8 hours, and then let it stand still. After the crystals sink completely, collect the crystals by centrifugation, wash the crystals once with 3% sodium chloride solution, and obtain wet crystals of sodium salt.

[0013] recrystallization

[0014] According to 26% dry analysis, according to dry weight 0.5 / 10 Calculate the total volume, add 1 gram of solid citric acid per gram of dry product,

[0015] Add water to dissolve to 1 / 3 of the total volume, let stand in the cold room for about 10 hours, filter, add 1.0 to the filtrate

[0016] Add 1 ml of 20% zinc acetate, 20 ml of cold acetone, and dilute with 5-10°C cold water to total

[0017] ...

Embodiment 2

[0020] Dissolve 1Kg of insulin salted-out product with 0.1N dilute hydrochloric acid to make a volume of 3 liters, adjust the pH to 7.8 with 10 equivalents of sodium hydroxide, add 0.15 liters of 30% sodium chloride solution, stir evenly, and then use 5N Adjust the pH to 7.8 with sodium hydroxide, stir at room temperature for 8 hours, and then let it stand still. After the crystals completely sink, collect the crystals by centrifugation, wash the crystals once with 3% sodium chloride solution, and obtain wet crystals of insulin sodium salt.

[0021] recrystallization

[0022] Dry by 30%, calculate total volume by dry weight 2 / 10, add 1.2 grams of solid citric acid by every gram of dry product,

[0023] Add water to dissolve to 1 / 3 of the total volume, let stand for about 15 hours, filter, add 1.5 ml of 20% zinc acetate and 20 ml of cold acetone to the filtrate according to 1 g of dry product, and dilute with cold water at 5-10 °C to For the total volume, adjust the pH...

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PUM

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Abstract

A process for purifying insuline includes such steps as salting out, sodium salt crystallizing, zinc salt recrystallizing, and collecting dried insuline crystal powder. Its advantages are short period and high output rate.

Description

technical field [0001] The present invention relates to a purification method of insulin, more specifically a crystallization method of insulin, which is used to prepare high-purity insulin. Background technique [0002] At present, most of the known insulin production adopts the zinc salt crystallization process, and the process flow is: salt solution solution → acid protein removal → filtration → zinc precipitation → dissolution → alkali protein removal → filtration → crystallization → collection and drying. Its disadvantages are low product yield, many operation steps, long production cycle, high consumption of auxiliary materials, and about 15% of recycled products are produced in the process at the same time, especially for the production of high-purity insulin. Obviously, its process is unreasonable. Chinese patent Publication No. CN1077203A discloses an invention patent application "a crystallization process of insulin sodium salt", the process of which is: salt solut...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C07K1/14C07K14/62
Inventor 罗桂荣彭淑华陈慧
Owner JIANGSU WANBANG BIOPHARMLS
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