Separation of oligosaccharides

A technology of oligosaccharides and neutral oligosaccharides, applied in the field of separation of neutral human milk oligosaccharides from each other, can solve problems such as not mentioned

Active Publication Date: 2021-08-06
GLYCOM AS
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Although this method provides 2'-FL with a purity of 94%-94.5% (by HPLC), the application makes no mention of the approximately 5% major carbohydrate contaminants present in the final product, such as 3-FL, DFL, and Lactose) is reduced by this method

Method used

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  • Separation of oligosaccharides
  • Separation of oligosaccharides

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Experimental program
Comparison scheme
Effect test

Embodiment 1

[0096] Two glass columns (1:40 cm, d: 26 mm) were filled with Purosorb PAD428 BPS-DVB resin and Sepabeads SP825L PS-DVB (non-brominated) resin, respectively. A solution of a mixture of LNnT (500 mg) and pLNnH (115 mg, ratio 4.35:1) dissolved in 10 ml of water was chromatographed on each resin with a flow rate of about 2.5 ml / min. Fractions (about 20ml) were collected and analyzed by HPLC.

[0097] result:

[0098] For Purosorb PAD428 brominated PS-DVB resin, fractions 3-10 (96 ml, about 0.5-1.67 bed volumes) were combined, which contained a ratio of LNnT:pLNnH of 8.7:1 (77% recovery of LNnT).

[0099] For Sepabeads SP825L PS-DVB resin, fractions 2-8 (104ml, about 0.3-1.17 bed volumes) were combined, which contained a ratio of LNnT:pLNnH of 4.7:1 (98% recovery of LNnT).

[0100] BPS-DVB resin enriched LNnT by doubling the ratio of LNnT, while unbrominated PS-DVB resin showed little separation.

Embodiment 2

[0102] A glass column (1:40 cm, d: 26 mm) was filled with Purosorb PAD428 BPS-DVB resin. A solution of a mixture of LNnT (1.2 g) and pLNnH (0.25 g, ratio 4.8:1) dissolved in 20 ml of water was subjected to chromatography with a flow rate of about 4 ml / min. Fractions (about 30ml) were collected and analyzed by HPLC.

[0103] Fractions 2-10 were pooled (240 ml, ca. 0.5-2.6 bed volumes), which contained LNnT:pLNnH in a ratio of 7.65:1 (90% recovery of LNnT).

Embodiment 3

[0105]A glass column (1:40 cm, d: 26 mm) was filled with Sepabeads SP207 BPS-DVB resin. LNnT (1.5 g), pLNnH (100 mg) and F-pLNnH (Galβ1-4[Fucα1-3]GlcNAcβ1-3Galβ1-4GlcNAcβ1-3Galβ1-4Glc, see WO 2016 / 063261, 50 mg; ratio 30:2:1) A solution of the mixture dissolved in 30 ml of water was subjected to chromatography with a flow rate of about 6 ml / min. Fractions (about 30ml) were collected and analyzed by HPLC.

[0106] Fractions 3-14 (400ml) were pooled and contained LNnT:pLNnH:Galβ1-4[Fucα1-3]GlcNAcβ1-3Galβ1-4GlcNAcβ1-3Galβ1-4Glc in a ratio of 129:2.6:1.

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Abstract

The invention relates to a method for the separation of two hydrophilic neutral oligosaccharides from each other with a chromatography on a bromine functionalized polystyrene cross-linked with divinylbenzene (BPS-DVB) stationary medium.

Description

technical field [0001] The present invention relates to a method for separating two hydrophilic neutral oligosaccharides, preferably at least two neutral human milk oligosaccharides (HMOs), produced by fermentation or enzymatic methods from each other. Specifically, one of at least two neutral oligosaccharides has a higher molecular weight than the other, and they show partitioning on the hydrophobic stationary phase. Background technique [0002] In recent years, increasing efforts have been made to produce industrially complex carbohydrates, such as secreted oligosaccharides. This is due to the role of this class of compounds in many biological processes in living organisms. Secreted oligosaccharides, such as human milk oligosaccharides (HMOs), have become particularly important commercial targets in nutritional and therapeutic applications. Human milk oligosaccharides have become of great interest in the past few years due to their important role in human development. ...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): B01D15/08C07H1/06C12P19/00
CPCC07H1/06C12P19/26C12P19/04C12P19/12B01D15/325B01J20/285B01D15/12B01D61/027B01J20/267B01D61/145Y02A50/30B01D2311/2697B01D15/08C07H3/06B01D15/125
Inventor N·汉任M·乔德利乌斯赫德罗斯P·沙萨捏
Owner GLYCOM AS
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