High Fidelity Restriction Endonucleases
US-2024352437-A1 · Oct 24, 2024 · US
Methods for purification of arylsulfatase A
US10336992B2 · US · B2
| Field | Value |
|---|---|
| Publication number | US-10336992-B2 |
| Application number | US-201214128468-A |
| Country | US |
| Kind code | B2 |
| Filing date | Jul 9, 2012 |
| Priority date | Jul 8, 2011 |
| Publication date | Jul 2, 2019 |
| Grant date | Jul 2, 2019 |
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Abstract
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Methods of producing arylsulfatase A are described herein. The methods can include one or more steps of chromatography. Exemplary chromatographic steps include ion exchange chromatography, mixed mode chromatography, and hydrophobic interaction chromatography. Compositions containing and methods using arylsulfatase A produced by the methods described herein are also disclosed.
First claim
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What is claimed: 1. A method of purifying recombinant arylsulfatase A (ASA) protein, comprising providing a sample of recombinant ASA protein; subjecting the sample to a first chromatography step which is anion exchange chromatography, subjecting the sample from the anion exchange chromatography to a second chromatography step which is mixed-mode chromatography, subjecting the sample from the mixed-mode chromatography to a third chromatography step which is hydrophobic interaction chromatography, and subjecting the sample from the hydrophobic interaction chromatography to a fourth chromatography step which is cation exchange chromatography, wherein fractions from each chromatography step containing recombinant ASA protein are determined by detecting the presence of recombinant ASA protein or enzymatic activity, and wherein the purified recombinant ASA protein has a specific activity of at least 75 U/mg, wherein 1 Unit of enzyme activity is defined as the hydrolysis of 1 μmol p-Nitrocatechol Sulphate (pNCS) per minute at 37° C., pH 5.0. 2. The method of claim 1 , wherein the sample of recombinant ASA protein is obtained from cultured cell extract or cell culture supernatant. 3. The method of claim 1 , wherein the cation exchange chromatography is sulfopropyl (SP) cation exchange chromatography. 4. The method of claim 1 , wherein the mixed-mode chromatography is hydroxyapatite (HA) chromatography. 5. The method of claim 1 , wherein the hydrophobic interaction chromatography is phenyl chromatography.
Assignees
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Classifications
- C12N9/16Primary
acting on ester bonds (3.1) · CPC title
Cerebroside-sulfatase (3.1.6.8) · CPC title
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- What does patent US10336992B2 cover?
- Methods of producing arylsulfatase A are described herein. The methods can include one or more steps of chromatography. Exemplary chromatographic steps include ion exchange chromatography, mixed mode chromatography, and hydrophobic interaction chromatography. Compositions containing and methods using arylsulfatase A produced by the methods described herein are also disclosed.
- Who is the assignee on this patent?
- Yang Ying, Wang Yong, Shire Human Genetic Therapies
- What technology area does this patent fall under?
- Primary CPC classification C12N9/16. Mapped technology areas include Chemistry & Metallurgy.
- When was this patent published?
- Publication date Tue Jul 02 2019 00:00:00 GMT+0000 (Coordinated Universal Time) (B2). Legal status and post-grant events are not shown on this page.
- What related patents are in patentsdb?
- We list 8 related publications on this page (citations in our corpus or others sharing the same primary CPC).