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Method for purifying transdermal peptide

A transdermal, separation and purification technology, applied in the field of purified transdermal peptides, can solve the problems of low purity and low purification efficiency of transdermal peptides, and achieve the effect of overcoming low purification efficiency

Pending Publication Date: 2020-02-04
安徽科门生物科技有限公司
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0004] In view of the deficiencies in the prior art, the purpose of the present invention is to provide a method for purifying transderma

Method used

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Examples

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Embodiment 1

[0019] A method for purifying transdermal peptides, the method comprising the following steps:

[0020] Step 1: Dissolve the crude transdermal peptide in the mixed solution to obtain a crude peptide solution, wherein the mixed solution is mixed with ether and pentane at a mass ratio of 1:2, the mass concentration of the mixed solution is 30%, and the crude transdermal peptide is mixed with The ratio of parts by weight of the solution is 1:20-30, and the dissolution conditions are: dissolve at 35-40°C for 0.5-0.8h, and filter after the dissolution to obtain the crude peptide solution;

[0021] Step 2: Take the crude peptide solution, and use C4 phase silica gel short column for gradient elution purification, wherein the C4 silica gel column size is: 5μm, 4.6×250mm, the mobile phase is two phases, mobile phase: A phase: yes volume Concentration is 0.25% formic acid aqueous solution, phase B: is acetonitrile solution with volume concentration of 0.25% formic acid, mobile phase gr...

Embodiment 2

[0025] A method for purifying transdermal peptides, the method comprising the following steps:

[0026] Step 1: Dissolve the crude transdermal peptide in the mixed solution to obtain a crude peptide solution, wherein the mixed solution is mixed with ether and pentane at a mass ratio of 1:2, the mass concentration of the mixed solution is 30%, and the crude transdermal peptide is mixed with The ratio of parts by weight of the solution is 1:20-30, and the dissolution conditions are: dissolve at 35-40°C for 0.5-0.8h, and filter after the dissolution to obtain the crude peptide solution;

[0027] Step 2: Take the crude peptide solution, and use C4 phase silica gel short column for gradient elution purification, wherein the C4 silica gel column size is: 5μm, 4.6×250mm, the mobile phase is two phases, mobile phase: A phase: yes volume The concentration is 0.4% formic acid aqueous solution, phase B: is the volume concentration of 0.4% formic acid acetonitrile solution, mobile phase g...

Embodiment 3

[0031] A method for purifying transdermal peptides, the method comprising the following steps:

[0032] Step 1: Dissolve the crude transdermal peptide in the mixed solution to obtain a crude peptide solution, wherein the mixed solution is mixed with ether and pentane at a mass ratio of 1:2, the mass concentration of the mixed solution is 30%, and the crude transdermal peptide is mixed with The ratio of parts by weight of the solution is 1:20-30, and the dissolution conditions are: dissolve at 35-40°C for 0.5-0.8h, and filter after the dissolution to obtain the crude peptide solution;

[0033] Step 2: Take the crude peptide solution, and use C4 phase silica gel short column for gradient elution purification, wherein the C4 silica gel column size is: 5μm, 4.6×250mm, the mobile phase is two phases, mobile phase: A phase: yes volume Concentration is 0.25% formic acid aqueous solution, phase B: is acetonitrile solution with volume concentration of 0.4% formic acid, mobile phase gra...

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Abstract

The invention discloses a method for purifying transdermal peptide. The method comprises the following steps: step 1, a crude transdermal peptide product is dissolved in a mixed solution to obtain a crude peptide solution, and filtration is performed, wherein the weight part ratio of the crude transdermal peptide product to the mixed solution being 1:(20-30); step 2, the crude peptide solution istaken and subjected to gradient elution and purification through a C4-phase silica gel short column, wherein a mobile phase comprises two phases including a phase A and a phase B, the phase A is a formic acid aqueous solution with the volume concentration being 0.25-0.4%, the phase B is a formic acid acetonitrile solution with the volume concentration being 0.25-0.4%, gradient elution, separationand purification are conducted, and a peptide solution with a target peak value is collected; and step 3, reduced pressure rotary evaporation concentration is performed on the separated and purified peptide solution with the purity of 99% or above, and a concentrated solution is collected to obtain purified transdermal peptide. The transdermal peptide with the purity of 99% is obtained by the method, the yield is 72% or above, and the problems of low purification efficiency and low purity of obtained transdermal peptide in a traditional method for purifying the transdermal peptide are solved.

Description

technical field [0001] The invention belongs to the field of purification of transdermal peptides, and in particular relates to a method for purifying transdermal peptides. Background technique [0002] Transdermal peptides are some small molecular polypeptides with the ability to penetrate cell membranes, which can effectively carry exogenous hydrophobic macromolecules that are twice their molecular weight into cells without significant toxic and side effects on host cells. There are multiple transmembrane mechanisms for transdermal peptides, such as direct penetration into the cell membrane, a temporary structure formed through translocation, and endocytosis-mediated membrane entry. The specific transmembrane mechanism is unclear, but it is generally believed that the direct contact between transdermal peptides and negatively charged substances on the cell surface is essential. The membrane-penetrating properties of transdermal peptides make them have broad application pr...

Claims

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Application Information

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IPC IPC(8): C07K1/20
CPCC07K1/20
Inventor 张鹏成
Owner 安徽科门生物科技有限公司
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