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Method for purifying polysaccharides

A technology of polysaccharides and neutral polysaccharides, applied in pharmaceutical formulations, medical preparations containing active ingredients, antibody medical ingredients, etc., can solve the burden of increasing manufacturing compliance, inefficiency, reduced yield and quality consistency, etc. question

Pending Publication Date: 2022-06-24
LONZA INC
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Additionally, the use of different reagents in each individual purification procedure can lead to less consistent yield and quality
In addition, the use of different processes can increase the burden of manufacturing compliance, as each step and reagent may need to be evaluated individually during process characterization, resulting in reduced efficiency and increased cost
Therefore, purification of polysaccharides can often be tedious, complex, time-consuming and inefficient work

Method used

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  • Method for purifying polysaccharides
  • Method for purifying polysaccharides
  • Method for purifying polysaccharides

Examples

Experimental program
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Effect test

example 1

[0099] The following procedure was tested to isolate polysaccharides from Streptococcus cells of two different serotypes (serotypes 3 and 4).

[0100] 1) Precipitate DNA with CTAB and NaCl;

[0101] 2) Depth filtration to remove sediment;

[0102] 3) Precipitation with KI to remove CTAB;

[0103] 4) Centrifuge to remove sediment;

[0104] 5) Carbon filtration to remove protein and other impurities;

[0105] 6) Ceramic hydroxyapatite type (CHT) silica gel column chromatography combined with SARTOBIND phenyl membrane chromatography to further remove impurities;

[0106] 7) Tangential flow filtration for polysaccharide concentration.

[0107] Reference cells were subjected to a solution containing CTAB and no NaCl. Test cells were subjected to a solution containing CTAB and 400 mM NaCl.

[0108] The total polysaccharide yields are described in Table 1:

[0109] Table 1.

[0110]

[0111] As shown in Table 1, cells contacted with solutions containing NaCl resulted in a ...

example 2

[0120] The following procedure is used to isolate polysaccharides, eg, positively charged, negatively charged and / or neutral polysaccharides (eg, pullulan), from A. pullulans cells. The cells are A. pullulans var. pullulan, A. pullulans var. pullulan, or both. The cells contain one of A. pullulans strain MC 571, strain MC 573, strain MC 574, strain MC576, strain MC 711, strain MC 737, strain MC 745, strain MC 767, strain CBS 701.76 and strain CBS 105.22. one or more.

[0121] 1) Precipitate DNA with CTAB and salt;

[0122] 2) Depth filtration to remove sediment;

[0123] 3) Precipitation with KI to remove CTAB;

[0124] 4) Centrifuge to remove sediment;

[0125] 5) Carbon filtration to remove protein and other impurities;

[0126] 6) Ceramic hydroxyapatite type (CHT) silica gel column chromatography combined with SARTOBIND phenyl membrane chromatography to further remove impurities;

[0127] 7) Tangential flow filtration for polysaccharide concentration.

[0128] The co...

example 3

[0130]The procedure described in Example 2 was used to isolate polysaccharides, eg, positively charged, negatively charged and / or neutral polysaccharides, from S. aureus cells. The cells contain one or more of S. aureus serotypes 1, 2, 5, 8, 336 or NT. The concentrations of CTAB used in the procedure are provided in Table 2. The concentrations and types of salts used in the procedure are provided in Table 3.

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Abstract

The present disclosure provides a method of purifying a polysaccharide from a cell lysate, the method comprising: partially purifying a cell lysate comprising impurities and a polysaccharide to obtain a clarified crude lysate; mixing the clarified crude lysate with a neutralizing solution comprising a salt to form a neutralized lysate; mixing the neutralized lysate with a precipitation solution comprising cetyltrimethylammonium bromide to form a first supernatant and a first precipitate; and separating the first precipitate from the first supernatant, wherein the polysaccharide is located in the first supernatant. The invention further provides a method for preparing the polysaccharide vaccine. Vaccines, delivery systems, compositions, and polysaccharides prepared by the methods described herein are also provided.

Description

technical field [0001] The present disclosure provides a method of purifying polysaccharides from cells and / or cell lysates. The present disclosure further provides a method for preparing a polysaccharide vaccine. Also provided are vaccines, delivery systems, compositions and polysaccharides prepared by the methods described herein. Background technique [0002] Pathogenic bacteria typically have cell surface capsules coated with polysaccharides. In a similar fashion, pathogenic fungi can have cell walls containing polysaccharides. After infection, these bacterial and fungal polysaccharides interfere with the immune response by preventing host antibodies from attaching to cells. These polysaccharides have been found to be effective vaccines against bacterial or fungal pathogens. In certain instances, multiple polysaccharides are conjugated into one vaccine formulation to increase protection against multiple serotypes or strains and / or improve the effectiveness of the vac...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): A61K39/09C08B37/00C12P19/04A61P31/04
CPCA61K39/092C08B37/0003A61P31/04C08L5/00C08L5/08A61K47/64A61K31/722A61K35/74A61K36/06A61K39/0002
Inventor A·祖尔布里根
Owner LONZA INC
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