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Method for preparing pegylation collagen and application thereof

A pegylated, collagen-like technology, applied in the biological field, can solve the problem of low strength of the bionic cornea, and achieve the effect of good cross-linking performance and high application prospects

Pending Publication Date: 2021-12-10
EOSVISION MEDTECH TECH CO LTD
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0008] In order to solve the problem of low strength of the existing bionic cornea, the invention provides a method for preparing pegylated collagen-like protein, the method comprising the following steps:

Method used

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  • Method for preparing pegylation collagen and application thereof
  • Method for preparing pegylation collagen and application thereof

Examples

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Embodiment 1-1

[0105] Example 1-1: A method for preparing pegylated collagen-like protein

[0106] The present embodiment provides a method for preparing pegylated collagen-like proteins, the method comprising the steps of:

[0107] Reaction steps: Collagen-like protein (the amino acid sequence of the collagen-like protein is shown in SEQ ID NO.1, and the N-terminal of the collagen-like protein is connected with a linker with an amino acid sequence shown in SEQ ID NO.2) and polyethylene glycol The derivative (8-arm-PEG-40k-MAL) is dissolved in the dilute hydrochloric acid solution whose concentration is 5mmol / L (the pH of the dilute hydrochloric acid solution is adjusted to 6.5 with the sodium hydroxide solution of pH 11.0), so that in the dilute hydrochloric acid solution, the The molar concentrations of collagen and polyethylene glycol derivatives were 2.4mmol / L and 0.3mmol / L respectively, and a reaction system was obtained; the reaction system was reacted for 12 hours at a pH of 6.5 and a...

Embodiment 2-1

[0109] Example 2-1: A method for preparing a freeze-dried preparation of pegylated collagen-like protein

[0110] This embodiment provides a method for preparing a freeze-dried preparation of pegylated collagen-like protein, the method comprising the following steps:

[0111] The PEGylated collagen-like protein 1 and mannitol prepared in Example 1 were mixed according to the mass ratio of 1:5 to obtain a lyophilized system; the lyophilized system was lyophilized to obtain a lyophilized preparation 1;

[0112] Wherein, lyophilization comprises the following stages:

[0113] Stage 1: Freeze-drying for 6 hours at a temperature of -45°C and a vacuum of 500m Torr;

[0114] Stage 2: Freeze-drying for 17 hours at a temperature of -30°C and a vacuum of 100 m Torr;

[0115] Stage 3: Freeze-drying for 7 hours at a temperature of 25° C. and a vacuum of 100 m Torr.

Embodiment 1-2

[0116] Embodiment 1-2: A kind of method for preparing pegylated collagen-like protein

[0117] This example provides a method for preparing PEGylated collagen-like proteins. On the basis of the method in Example 1-1, the reaction temperature (5°C) is replaced by: 2°C, 8°C, At 25°C and 37°C, PEGylated collagen-like proteins 2-5 were prepared.

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Abstract

The invention relates to a method for preparing pegylation collagen and application thereof, and belongs to the technical field of biology. The method for preparing pegylation collagen comprises the step that collagen and polyethylene glycol derivatives react under the conditions that the pH is 6.0-8.0 and the temperature is 2-8 DEG C to prepare pegylation collagen. Compared with other existing methods for preparing pegylation collagen, pegylation collagen prepared through the method is better in crosslinking performance, and after the pegylation collagen prepared through the method is prepared into hydrogel to form a cornea, the strength can be improved by 10 times or above compared with pegylation collagen prepared through the existing methods; and in addition, the reaction time for preparing pegylation collagen through the method is shorter, and pegylation collagen capable of being prepared into hydrogel to form the cornea can be obtained only through the reaction of 5-8h.

Description

technical field [0001] The invention relates to a method for preparing pegylated collagen-like protein and its application, belonging to the field of biotechnology. Background technique [0002] The cornea is the convex, highly transparent substance at the front of the eye. It is in the shape of a transverse ellipse, covering the iris, pupil and anterior chamber, and provides most of the refractive power for the eye. Coupled with the refractive power of the lens, the light can be accurately focused on the retina to form an image. The cornea has very sensitive nerve endings. If something foreign comes into contact with the cornea, the eyelids will involuntarily close to protect the eyes. In order to maintain transparency, the cornea does not have blood vessels, and obtains nutrients and oxygen through tears and aqueous humor. [0003] The cornea is very fragile, eye trauma, inflammation, allergic reaction, physical damage, chemical burns, strenuous exercise, overuse of the ...

Claims

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Application Information

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IPC IPC(8): C07K14/78C07K1/107C07K1/34A61L27/24A61L27/52
CPCC07K14/78A61L27/24A61L27/52A61L2430/16C08L89/00
Inventor 王崇宇张加慧张军刘沐荣何超先
Owner EOSVISION MEDTECH TECH CO LTD
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