Method for purifying glycopeptide compound

A purification method and compound technology, which is applied in the field of glycopeptide compounds, can solve the problems of slow desorption speed, large difference in swelling coefficient, tailing, etc., and achieve the effects of reducing yield loss, shortening purification cycle, and improving quality

Active Publication Date: 2011-05-11
SHANGHAI LAIYI BIOMEDICAL RES & DEV CENT
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, dextran gel has defects such as large difference in swelling coefficient in different solvents, low resolution, severe tailing during desorption, and too slow desorption speed, which is not an ideal preparation process.

Method used

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  • Method for purifying glycopeptide compound
  • Method for purifying glycopeptide compound
  • Method for purifying glycopeptide compound

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0027] Example 1 , Compound 3 preparation and crude purity

[0028] 1.1. Preparation

[0029] Referring to the fermentation method in the Chinese Invention Patent Application No. 200910053906.9, the fermentation broth of compound 3 was prepared, and the titer was 0.3 mg / ml after testing.

[0030] 1.2, crude and pure

[0031] Take 20L of the fermentation broth prepared in step 1.1, adjust the pH to 9.0 and filter, load the obtained clarified filtrate into 500ml of macroporous adsorption resin XAD-1600 (Rohm & Haas), and then wash the resin with 8% ethanol aqueous solution.

[0032] Then eluted with 2L 0.05% HCl aqueous solution, collected the eluate, concentrated to 500ml with a nanofiltration membrane with a 300Da pore size, and then added 100g of gel-type cation exchange resin 001×4 for static decolorization for 6 hours.

[0033] The resin was filtered off, and the decolorized solution was lyophilized to obtain 4.3 g of a crude product of compound 3.

[0034] After testin...

Embodiment 2-10

[0035] Examples 2-10 , Compounds 1, 2, 4-8 are pure

[0036] Take 0.20 g of each of the crude products of compounds 1, 2, 4-8 shown in Table 2, dissolve them in 10 ml of pure water, and load the samples onto a 10 mm × 20 cm glass chromatography column equipped with 10 ml of reversed-phase polymer filler Uni PS. , the sample flow rate was 10 ml / h. After sample loading, pre-wash with 10 ml methanol aqueous solution for 1 h, and then elute with methanol aqueous solution with or without HCl at a flow rate of 20 ml / h. After elution of 6ml, the eluate was collected, and a total of 15ml of eluate was collected. The eluate was concentrated and dried to obtain the purified products of compounds 1, 2, 4-8, respectively. The chromatographic purity of the crude product and the middle of the purification step The used pre-wash solution and elution solution are shown in Table 2, respectively. The purified products of the above compounds obtained were detected and the yields of the above ...

Embodiment 11-19

[0043] Examples 11-19 , Compound 3 is pure

[0044] Take 6 parts of the crude product of compound 3 obtained in Example 1, each 0.20 g, dissolve in 10 ml of pure water, respectively, and then load the samples into a 10 mm × 20 cm glass chromatography column equipped with 10 ml of reversed-phase polymer filler Uni PS. , after the sample loading, pre-wash with 10ml methanol aqueous solution for 1h, and then elute with methanol aqueous solution with or without HCl. After eluting 6ml, start to collect the eluent, and collect a total of 15ml of eluent, and then eluate the eluate Concentrate and dry to obtain the purified product of compound 3, the purification step parameters are shown in Table 4, and the obtained purified product of compound 3 is detected, and the detection results are shown in Table 5.

[0045] Table 4. Refinement parameters

[0046]

[0047] Table 5, the detection result of purified product

[0048] Example

Yield

color

purity

yiel...

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Abstract

The invention provides a method for purifying a glycopeptide compound, which comprises the following steps: A) loading a sample of aqueous solution of a glycopeptide compound onto an inverse polymer filler Uni PS; B) pre-washing by using pre-washing solution; and C) eluting with eluting solution. When the method provided by the invention is used, the quality of target components of the glycopeptide compound can be improved effectively, the purification period is shortened, and the yield loss is reduced. In addition, the method can be used for preparing a small amount of sample of the glycopeptide compound in a laboratory, and also can be used for industrial production of the glycopeptide compound.

Description

technical field [0001] The invention belongs to the field of glycopeptide compounds, and in particular relates to a purification method for glycopeptide compounds. Background technique [0002] Existing glycopeptide compounds generally use reversed-phase silica gel or dextran gel as a medium for purification and preparation. Among them, the most widely used reversed-phase silica gel is C18. In aqueous solution), a water-soluble organic solvent aqueous solution (or an aqueous solution containing a buffer salt) is used as the eluent, and the desorbed liquid is subjected to post-treatment such as desalting and drying to obtain a pure product. Due to the disadvantages of high cost, low yield, serious irreversible adsorption, and poor medium tolerance, this method can generally only be used for the preparation of a small number of samples in the laboratory. [0003] In industrial production, Sephadex is used as a medium for purification and preparation. For example, G-15 is used...

Claims

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Application Information

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IPC IPC(8): C07K1/20C07K9/00
Inventor 阮林高朱丽夏兴李秋爽陈代杰戈梅
Owner SHANGHAI LAIYI BIOMEDICAL RES & DEV CENT
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