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Method for preparing beta-amyloid polypeptide 1-42 pure product

A technology for amyloid peptides and pure products, which is applied in the field of high performance liquid chromatography to prepare pure products of β-amyloid peptides 1-42, which can solve problems such as high equipment requirements and cumbersome mobile phase systems, and achieve cost-saving effects

Active Publication Date: 2010-03-03
GL BIOCHEM SHANGHAI
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

To solve the technical problems of high equipment requirements and cumbersome mobile phase system used in the preparation of existing high performance liquid chromatography methods

Method used

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  • Method for preparing beta-amyloid polypeptide 1-42 pure product
  • Method for preparing beta-amyloid polypeptide 1-42 pure product
  • Method for preparing beta-amyloid polypeptide 1-42 pure product

Examples

Experimental program
Comparison scheme
Effect test

specific Embodiment approach

[0029] Specific embodiments: dissolving of the crude product of β-amyloid polypeptide 1-42, filtration: take 100-200mg of crude product of β-amyloid polypeptide 1-42 and dissolve it in 10ml of acetonitrile aqueous solution, and the acetonitrile aqueous solution is composed of 30-50% acetonitrile and 70- Mixing with 50% water, ultrasonic treatment, until the sample is completely clarified, filter with a 0.45 μ organic phase filter membrane (Shanghai Yadong Nuclear Grade Resin Co., Ltd.), and take the filtrate for later use.

[0030] Chromatographic conditions:

[0031] LC3000 preparative high liquid chromatograph (high pressure double pump, UV detector) and CXTH-3000 chromatographic workstation of Beijing Innovation Tongheng Technology Co., Ltd. Mobile phase: A: an aqueous solution with a concentration of 0.1-0.2% trifluoroacetic acid, B: an acetonitrile solution with a concentration of 0.1-0.2% trifluoroacetic acid, 0 to 25 minutes A: B from 90-70: 10-30 to 10- 30:90-70; Flow...

Embodiment 1

[0043] Example 1: The loading amount is 100mg, mobile phase: A: concentration is the aqueous solution of 0.1% trifluoroacetic acid, B: the concentration is the acetonitrile solution of 0.1% trifluoroacetic acid, the sample dissolution condition is volume ratio: acetonitrile: water=30 : 70, 0 to 25 minutes A: B from 90: 10 to 10: 90, and the rest are the same as above.

Embodiment 2

[0044] Example 2, sample loading is 150 mg, mobile phase: A: aqueous solution with concentration of 0.15% trifluoroacetic acid, B: acetonitrile solution with concentration of 0.15% trifluoroacetic acid, sample dissolution condition is volume ratio: acetonitrile: water=40 : 60, 0 to 25 minutes A: B from 80: 20 to 20: 80, and the rest are the same as above.

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Abstract

The invention relates to a polypeptide preparation method, in particular to a method for using high-efficiency liquid chromatography to prepare beta-amyloid 1-42 pure product, to resolve the technicalproblems of prior high-efficiency liquid chromatography as high equipment demands and complex flowing system. The preparation conditions comprise that PLRP-S, 10-15um, 21.2x250mm(I.D), flowing phase:A: 0.1-0.2% trifluoroacetic acid water, B: 0.1-0.2% trifluoroacetic acid acetonitrile, 0-25min, A:B is from 90-70:10-30 to 10-30:90-70, flowing speed is 15ml / min, wavelength is 220nm, sample densityis 100-200mg / 10ml and sample amount is 100-200mg. The preparation method uses general agent of high-efficiency liquid chromatography as flowing phase and eliminates other machines beside high-efficiency liquid chromatography machine, to save cost and simply obtain pure product quickly.

Description

Technical field: [0001] The invention relates to a preparation method of polypeptide, in particular to a method for preparing pure β-amyloid polypeptide 1-42 by high performance liquid chromatography. Background technique: [0002] Alzheimer's disease (AD) is a degenerative disease of the central nervous system with a high incidence and great harm. Nodules, the number of neurons mainly in the hippocampus and cortical areas decreased. β-amyloid polypeptide 1-42 (Amyloidβ1-42peptide, referred to as Aβ42) is a specific pathological change in the brain tissue of patients with Alzheimer's disease, and it is also changed in the cerebral effusion and plasma of AD patients . Studies have shown that the deposition of Amyloidβ1-42peptide in the brain is the main cause of Alzheimer's disease, so pure amyloidβ1-42peptide plays a very important role in the pathological research of Alzheimer's disease. [0003] The preparation method report of Amyloid β1-42peptide crude product is seld...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C07K1/16
Inventor 徐红岩任丽萍李冬冬
Owner GL BIOCHEM SHANGHAI