Separation method of sialoglycopeptide

A technology for the separation and separation of sialic acid sugars, applied in the field of separation and purification of glycopeptide compounds, which can solve the problems of short cycle, low cost, and many steps, and achieve the effects of stable purification method, shortened purification cycle, and improved purification efficiency

Inactive Publication Date: 2012-03-21
SHANDONG UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0007] Aiming at the shortcomings of many separation steps, long period and high cost in the separation method in the prior art, the purpose of the present invention is to provide a separation and purification method of sialic acid glycopeptide with short period, low cost and high efficiency

Method used

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  • Separation method of sialoglycopeptide
  • Separation method of sialoglycopeptide

Examples

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Embodiment 1

[0038] 1. Egg yolk pretreatment

[0039] First take 100 fresh unfertilized egg yolks (1.9 L), add an equal volume of deionized water to dilute, then add 1 / 5 volume (380 ml) of phenol / water (phenol:water volume ratio 9:1), 4 °C stirring was continued for 3 hours. Add 4.8 L of deionized water to the mixture, mix well and centrifuge (10000 g, 25 minutes), and concentrate the supernatant to 50 ml with a rotary evaporator to obtain a sample containing sialic acid glycopeptide.

[0040] 2. Sephadex G-50 separation

[0041] The Sephadex G-50 column (1.6cm×100cm) was connected with a fast liquid chromatograph, using 0.1M NaCl as the eluent, the flow rate: 1ml / min, and the ultraviolet detection wavelength: 214nm. Place the column in an incubator at a temperature of 25°C. Add 2ml of the sample described in step 1, and collect the 110-140ml sample of the eluent, such as figure 1 As shown, collect the sample marked "Collection Area".

[0042] 3. Concentrate the eluate and desalt and ...

Embodiment 2

[0053] 1. Egg yolk pretreatment

[0054] First take 100 fresh unfertilized egg yolks (1.9L), add an equal volume of deionized water to dilute, then add 1 / 5 volume (380ml) of phenol / water (volume ratio 9:1), and keep stirring at 4°C for 3 Hour. Add 4.8 L of deionized water to the mixture, mix well and centrifuge (10000 g, 25 minutes), and concentrate the supernatant to 50 ml with a rotary evaporator to obtain a sample containing sialic acid glycopeptide.

[0055] 2. Sephadex G-50 separation

[0056] The Sephadex G-50 column (1.6cm×100cm) was connected with a fast liquid chromatograph, using 0.1M NaCl as the eluent, the flow rate: 1ml / min, and the ultraviolet detection wavelength: 214nm. Place the column in an incubator at a temperature of 23°C. Add 2.5ml of the sample described in 1 above, and collect the 110-140ml sample of the eluate.

[0057] 3. Concentrate the eluate and desalt and purify it with an activated carbon column

[0058] Put 50 ml of the eluent into a 100 ml...

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Abstract

The invention discloses a separation method of sialoglycopeptide, including the following steps: (1) taking a fresh egg yolk, treating the fresh egg yolk with phenol and then conducting rotation evaporation to obtain a sample containing the sialoglycopeptide; (2) separating the sample with sephadex G-50 and collecting eluent; (3) concentrating the eluent, absorbing the concentrated sample with active carbon and then eluting the sample in acetonitrile solution to obtain eluent containing the sialoglycopeptide; and (4) freezing and drying the eluent to obtain pure sialoglycopeptide. In the method, the steps of G-25 desalting, HPLC (High Performance Liquid Chromatography) purification, ion exchange chromatography purification and the like of the existing method are eliminated, therefore the purification period is shortened greatly, the efficiency is improved, the cost is saved and inspected by HPLC, and the purity of the prepared sialoglycopeptide can achieve 99%. The separation method is used for not only laboratories but also small-scale industrial production.

Description

technical field [0001] The present invention relates to a method for separation and purification of glycopeptide compounds, in particular to a method for separation and purification of glycopeptides with natural complex N sugar chains such as sialic acid glycopeptides. Background technique [0002] The structure of sialylglycopeptides is: [0003] [0004] Sialylglycopeptides are glycopeptides with a complete natural complex N-glycan chain, with a relative molecular mass of 2866. Although a variety of oligosaccharides can be synthesized by chemical methods, the current technology still cannot synthesize this complete sialylated N-glycoside. Therefore, this sialoglycopeptide can only be isolated and purified from natural products. [0005] At present, the international method for separating and purifying sialoglycopeptide from egg yolk is mainly based on the purification method reported by Akira Seko of Japan in 1997. In this method, the egg yolk is firstly treated, sep...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C07K9/00C07K1/36C07K1/16
Inventor 陈敏邹洋吴志刚王鹏
Owner SHANDONG UNIV
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