Method of purifying polysaccharides
AssigneeLonza Ltd.
InventorAndreas Zurbriggen
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Abstract
The present disclosure provides a method of purifying polysaccharides from a cell lysate, comprising partially purifying the cell lysate comprising an impurity and a polysaccharide to obtain a clarified crude lysate; mixing the clarified crude lysate with a neutralization 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 present disclosure further provides a method of making a polysaccharide vaccine. Also provided are vaccines, delivery systems, compositions and polysaccharides made by the methods described herein.
Claims22
A method of purifying polysaccharides from a cell lysate, the method comprising
- a. partially purifying the cell lysate comprising a polysaccharide and an impurity to obtain a clarified crude lysate;
- b. mixing the clarified crude lysate with a neutralization solution to form a neutralized lysate comprising about 100 mM to about 2 M salt;
- c. mixing the neutralized lysate with a precipitation solution comprising cetyltrimethylammonium bromide (CTAB) to form a first supernatant and a first precipitate, wherein the first supernatant comprises about 0.5% v/v to about 4% v/v CTAB;
- and
- d. separating the first precipitate from the first supernatant,
- wherein the polysaccharide is substantially located in the first supernatant;
- and
- wherein the method is capable of purifying positively charged, negatively charged, and neutral polysaccharides, and wherein the positively charged, negatively charged, and neutral polysaccharides are substantially located in the first supernatant.
A method of purifying polysaccharides from a cell lysate, the method comprising
- a. partially purifying the cell lysate comprising an impurity and a polysaccharide to obtain a clarified crude lysate;
- b. mixing the clarified crude lysate with a neutralization solution to form a neutralized lysate comprising about 400 mM sodium chloride (NaCl);
- c. mixing the neutralized lysate with cetyltrimethylammonium bromide (CTAB) to form a first supernatant and a first precipitate, wherein the first supernatant comprises about 1.0% v/v CTAB;
- d. separating the first precipitate from the first supernatant;
- e. mixing the first supernatant with potassium iodide (KI) to form a second precipitate and a second supernatant;
- and
- f. separating the second precipitate from the second supernatant,
- wherein the polysaccharide is substantially located in the second supernatant;
- and
- wherein the method is capable of purifying positively charged, negatively charged, and neutral polysaccharides, and wherein the positively charged, negatively charged, and neutral polysaccharides are substantially located in the first supernatant.
Dependencies are inferred from the claim text.