Looking for breakthrough ideas for innovation challenges? Try Patsnap Eureka!

Method for purifying thymosin beta4

A thymosin and chromatography technology, applied in the field of batch industrialized purification of thymosin β4 (Thymosin β4), can solve the problems of low purity of thymosin β4, and achieve the effects of low cost, fast efficiency and high yield

Inactive Publication Date: 2012-09-19
滨海吉尔多肽有限公司 +2
View PDF3 Cites 5 Cited by
  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0006] The purpose of the present invention is to provide a method suitable for industrialized pure thymosin β4, which mainly solves the technical problem that the purity of thymosin β4 obtained by separation in the prior art is relatively low

Method used

the structure of the environmentally friendly knitted fabric provided by the present invention; figure 2 Flow chart of the yarn wrapping machine for environmentally friendly knitted fabrics and storage devices; image 3 Is the parameter map of the yarn covering machine
View more

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0017] 1. Sample treatment: Dissolve the synthesized thymosin β4 in deionized water (dissolution concentration is about 50mg / ml), filter through a filter membrane with a pore size of 0.45um, and collect the filtrate for later use.

[0018] 2. purification

[0019] Purification conditions: Chromatographic column: a chromatographic column with tetraalkyl silica gel bonded silica gel as the stationary phase, the column diameter and length: 5cm×25cm. Mobile phase: A phase: aqueous ammonium dihydrogen phosphate solution, adjust the pH with analytically pure phosphoric acid solution To 3.0~4.8; Phase B: chromatographically pure acetonitrile. Flow rate: 40-55ml / min. Detection wavelength: 220nm. Gradient: B%: 20-60%, 30-50min. The injection volume is 1.1-1.8g;

[0020] Purification process: wash the chromatographic column with acetonitrile with a concentration of more than 60% by mass, and then load the sample, and the sample volume is the sample solution. Linear gradient el...

Embodiment 2

[0024] 1. Sample treatment: Dissolve the synthesized thymosin β4 in deionized water (dissolution concentration is about 50mg / ml), filter through a filter membrane with a pore size of 0.45um, and collect the filtrate for later use.

[0025] 2. purification:

[0026] Purification conditions: Chromatographic column: a chromatographic column with octadecyl silica gel bonded silica gel as the stationary phase, column diameter and length: 5cm×25cm. Mobile phase: A phase: aqueous ammonium dihydrogen phosphate solution adjusted with analytically pure phosphoric acid solution pH to 3.5~5.3; Phase B: chromatographically pure acetonitrile. Flow rate: 40-55ml / min. Detection wavelength: 220nm. Gradient: B%: 23-65%, 40-60min. The injection volume is 1.0-1.7g;

[0027] Purification process: wash the chromatographic column with acetonitrile with a concentration of more than 60% by mass, and then load the sample, and the sample volume is the sample solution. Linear gradient elution, col...

Embodiment 3

[0031] 1. Sample treatment: Dissolve the synthesized thymosin β4 in deionized water (dissolution concentration is about 50mg / ml), filter through a filter membrane with a pore size of 0.45um, and collect the filtrate for later use.

[0032] 2. purification:

[0033] Purification conditions: Chromatographic column: a chromatographic column with octadecyl silica gel bonded silica gel as the stationary phase, column diameter and length: 10cm×50cm. Mobile phase: A phase: aqueous ammonium dihydrogen phosphate solution adjusted with analytically pure phosphoric acid solution pH to 3.5~4.8; Phase B: chromatographically pure acetonitrile. Flow rate: 60-80ml / min. Detection wavelength: 220nm. Gradient: B%: 20-65%, 60-90min. The injection volume is 8.0-10.0g;

[0034] Purification process: Rinse the chromatographic column with more than 60% acetonitrile and load the sample. The sample volume is the sample solution. Linear gradient elution, collect the target peak, and put the colle...

the structure of the environmentally friendly knitted fabric provided by the present invention; figure 2 Flow chart of the yarn wrapping machine for environmentally friendly knitted fabrics and storage devices; image 3 Is the parameter map of the yarn covering machine
Login to View More

PUM

No PUM Login to View More

Abstract

The invention discloses a method for purifying thymosin beta4 with a reversed phase high performance liquid chromatography, which is mainly used for solving the technical problem of low purity of thymosin beta4 obtained by separating in the prior art. According to the technical scheme of the invention, the method comprises the following steps of: (1) dissolving a crude peptide obtained by synthesizing solids with deionized water, gradually eluting and purifying by taking a reversed phase silica gel column of which the immobile phase is tetraalkylsilane bonded silica gel or octadecyl bonded silica gel and the mobile phase is a phosphate buffer solution as a phase A and taking chromatographically-pure acetonitrile as a phase B, and collecting a peptide solution of a target peak value; (2) switching and transforming into acetate by using an HPLC (High Performance Liquid Chromatography) method; and (3) and performing spinning evaporation, concentration and freeze drying on a final high-purity peptide solution under reduced pressure to obtain a powdery finished peptide. The invention provides a method for purifying thymosin beta4, which is suitable for industrialization. According to the method, fine peptides of which the purities are over 98.0 percent can be obtained, batch production is available, and the requirements of high purity, high yield, low cost and high efficiency are met.

Description

technical field [0001] The invention belongs to the technical field of reversed-phase HPLC, and in particular relates to a method for industrially purifying thymosin β4 (Thymosin β4) in batches. Background technique [0002] Thymosin β4 is a small peptide substance isolated from the thymus in the 1980s and found to be widely present in many tissues. It is an important actin-binding protein, containing 43 amino acids, with a molecular weight of about 5kD, with an isoelectric point of 5.1, which is highly conserved in mammals. Thymosin β 4 is a small molecular protein composed of 43 amino acids, which has a strong epithelial cell stimulation and regeneration effect, and plays an important role in embryonic heart development. Thymosin-β4 (thymosin-β4, Tb4) was originally isolated and purified from calf thymus fraction Ⅴ. Thymosin β4 was found to exist in almost all somatic cells except red blood cells, with the highest content in platelets, and also distributed in plasma and ...

Claims

the structure of the environmentally friendly knitted fabric provided by the present invention; figure 2 Flow chart of the yarn wrapping machine for environmentally friendly knitted fabrics and storage devices; image 3 Is the parameter map of the yarn covering machine
Login to View More

Application Information

Patent Timeline
no application Login to View More
IPC IPC(8): C07K14/575C07K1/20
Inventor 徐红岩闫凤张宏伟
Owner 滨海吉尔多肽有限公司
Who we serve
  • R&D Engineer
  • R&D Manager
  • IP Professional
Why Patsnap Eureka
  • Industry Leading Data Capabilities
  • Powerful AI technology
  • Patent DNA Extraction
Social media
Patsnap Eureka Blog
Learn More
PatSnap group products

Browse by: Latest US Patents, China's latest patents, Technical Efficacy Thesaurus, Application Domain, Technology Topic, Popular Technical Reports.

© 2024 PatSnap. All rights reserved.Legal|Privacy policy|Modern Slavery Act Transparency Statement|Sitemap|About US| Contact US: help@patsnap.com