Method for removing heat source from recombinant IL-18 and prepared product

A heat source and mobile phase technology, applied in the biological field, can solve the problems of strict ultrafiltration conditions, limited effect of removing heat source, and easy to be polluted.

Inactive Publication Date: 2020-10-27
HE EYE HOSPITAL SHENYANG +1
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

The activated carbon method will also adsorb the sample while adsorbing the heat source, reducing the sample concentration; the acid-base method will destroy the activity of the biological preparation itself; Pollution; most of the ion exchange method and affinity chromatography cannot adopt a process to remove endotoxin up to the standard
The above methods have limited effect on removing heat source and are not suitable for all samples

Method used

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  • Method for removing heat source from recombinant IL-18 and prepared product
  • Method for removing heat source from recombinant IL-18 and prepared product
  • Method for removing heat source from recombinant IL-18 and prepared product

Examples

Experimental program
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Effect test

Embodiment 1

[0036] 1. Purification of recombinant IL-18 by preparative liquid phase method

[0037] The prepared liquid is used to purify and remove endotoxin relative to recombinant IL-18. The sample is the liquid sample collected from the gel chromatography column of recombinant IL-18 prepared by Patent No.: 201510455749.X.

[0038] The chromatographic column medium is C18, the specification is 21.2mm×250mm, and the particle size is 7μm.

[0039]Mobile phase A: a mixed solution of pyrogen-free water and 0.1% trifluoroacetic acid, for example: add 1ml trifluoroacetic acid to 1L pyrogen-free water.

[0040] Mobile phase B: a mixed solution of acetonitrile and 0.1% trifluoroacetic acid, for example, add 1ml trifluoroacetic acid to 1L acetonitrile.

[0041] The mobile phase A and mobile phase B were used as eluents, the flow rate was 10 mL / min, the gradient elution mode was used for elution, the column temperature was 25°C, and the detection wavelength was 280 nm.

[0042] Gradient elutio...

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Abstract

The invention relates to the technical field of biology and particularly relates to a method for removing a heat source from recombinant IL-18 and a prepared product. According to the purification method provided by the invention, the recombinant IL-18 is obtained through carrying out purification by employing high-performance liquid chromatography, and chromatographic conditions are reasonable, so that good separation of the recombinant IL-18 from the heat source is achieved; and through recovering chromatographic peaks of the recombinant IL-18, the heat-source-removed recombinant IL-18 in line with requirements of articles for injection is obtained. Shown by experiments, the heat-source-removed recombinant IL-18 obtained through carrying out purification by employing the method providedby the invention has a concentration of 15.1mg / ml, the recovery rate is 85.32%, the heat source content is satisfactory, and physiological activity of the heat-source-removed recombinant IL-18 is notaffected.

Description

technical field [0001] The invention relates to the field of biotechnology, in particular to a method for removing heat source in recombinant IL-18 and a product obtained therefrom. Background technique [0002] Diabetic fundus vascular disease (DR) and age-related macular degeneration (AMD) are the main factors affecting vision and even blindness in advanced diabetes and the elderly. Since DR and AMD are a long-term pathological process, the abnormal growth of new blood vessels and metabolites will block the subretina, so that important nutrients cannot enter the retina from the blood, and the discharge of waste will also be obstructed, which will cause retinal cell damage. death, eventually leading to permanent blindness. Therefore, promoting the maturation of blood vessels while inhibiting the growth of blood vessels, so that more nutrients can enter the retina and remove existing metabolites, is the fundamental goal of DR and AMD treatment. [0003] In recent years, st...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C07K14/54C07K1/20A61K38/20A61P27/02A61P35/00
CPCA61K38/20A61P27/02A61P35/00C07K14/54
Inventor 张朔李雪娇冯铃刘辉赵航吴骁刘畅赵洪礼何向东何伟
Owner HE EYE HOSPITAL SHENYANG
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