Title:
PREPARATION METHOD FOR POLYPEPTIDE
Document Type and Number:
WIPO Patent Application WO/2022/241831
Kind Code:
A1
Abstract:
A preparation method for a polypeptide, the preparation method comprising the following steps: constructing an engineering strain of a Sumo tag fusion expression polypeptide gene, and inducing the engineering strain for the soluble expression of a polypeptide; purifying a crude protein containing a polypeptide precursor from the engineering strain; digesting the crude protein containing the polypeptide precursor by using Ulp1 protease to remove the Sumo tag; purifying a cleavage product of the Ulp1 protease by means of acetonitrile combined with heating precipitation or by using a method for hexafluoroisopropanol precipitation, so as to obtain the polypeptide. Efficient technology based on the soluble recombinant expression of a medicinal polypeptide or a precursor thereof has been established, a simple purification process based on one-step precipitation purification has been established, and further in combination with HPLC purification, a polypeptide purity of more than 97% can be achieved.
Inventors:
HONG HAO (US)
JAMES GAGE (US)
XIAO YI (CN)
ZHANG NA (CN)
JIAO XUECHENG (CN)
ZHAO JUNQI (CN)
WANG LEI (CN)
MENG XIANGYU (CN)
ZHANG MUJIAO (CN)
JAMES GAGE (US)
XIAO YI (CN)
ZHANG NA (CN)
JIAO XUECHENG (CN)
ZHAO JUNQI (CN)
WANG LEI (CN)
MENG XIANGYU (CN)
ZHANG MUJIAO (CN)
Application Number:
PCT/CN2021/097429
Publication Date:
November 24, 2022
Filing Date:
May 31, 2021
Export Citation:
Assignee:
ASYMCHEM LAB TIANJIN CO LTD (CN)
International Classes:
C12P21/02; C12P21/06
Foreign References:
| CN101413009A | 2009-04-22 | |||
| CN112142848A | 2020-12-29 | |||
| CN105647960A | 2016-06-08 | |||
| CN111269308A | 2020-06-12 | |||
| US20030086918A1 | 2003-05-08 |
Other References:
"Food Ingredients and Food Additive Analyzing Method", 31 August 2008, CHINA LIGHT INDUSTRY PRESS, CN, ISBN: 978-7-5019-6467-3, article OTLES S.: "4.2.2 protein removing process", pages: 42 - 44, XP009541318
TIAN YING, YUAN CAI-YUN, XU JING, LU GUANG-MING, LING ZHENG: "Small ubiquitin-related modifier fusion expression and characterization of human soluble B cell activating factor belonging to the TNF family in E. coli", YIXUE YANJIUSHENG XUEBAO [JOURNAL OF MEDICAL POSTGRADUATES], vol. 26, no. 11, 30 November 2013 (2013-11-30), pages 1147 - 1151, XP093006193, ISSN: 1008-8199, DOI: 10.16571/j.cnki.1008-8199.2013.11.008
XIE QIUBO, CHANG QIUSHUANG; ZHOU PENG; CUI LIWEI; GUO LING; LIU WEI; ZHANG QINGHUA; LU GENSHENG: "Soluble expression of murine interferon-epsilon in Escherichia coli", IMMUNOLOGICAL JOURNAL, DI-3 JUNYI DAXUE, CHINA, vol. 33, no. 3, 31 March 2017 (2017-03-31), China , pages 245 - 250, XP093006195, ISSN: 1000-8861, DOI: 10.13431/j.cnki.immunol.j.20170044
GUERRERO FERNANDO, CIRAGAN ANNIKA, IWAÏ HIDEO: "Tandem SUMO fusion vectors for improving soluble protein expression and purification", PROTEIN EXPRESSION AND PURIFICATION, ACADEMIC PRESS, SAN DIEGO, CA., vol. 116, 1 December 2015 (2015-12-01), SAN DIEGO, CA. , pages 42 - 49, XP093006196, ISSN: 1046-5928, DOI: 10.1016/j.pep.2015.08.019
TIAN YING, YUAN CAI-YUN, XU JING, LU GUANG-MING, LING ZHENG: "Small ubiquitin-related modifier fusion expression and characterization of human soluble B cell activating factor belonging to the TNF family in E. coli", YIXUE YANJIUSHENG XUEBAO [JOURNAL OF MEDICAL POSTGRADUATES], vol. 26, no. 11, 30 November 2013 (2013-11-30), pages 1147 - 1151, XP093006193, ISSN: 1008-8199, DOI: 10.16571/j.cnki.1008-8199.2013.11.008
XIE QIUBO, CHANG QIUSHUANG; ZHOU PENG; CUI LIWEI; GUO LING; LIU WEI; ZHANG QINGHUA; LU GENSHENG: "Soluble expression of murine interferon-epsilon in Escherichia coli", IMMUNOLOGICAL JOURNAL, DI-3 JUNYI DAXUE, CHINA, vol. 33, no. 3, 31 March 2017 (2017-03-31), China , pages 245 - 250, XP093006195, ISSN: 1000-8861, DOI: 10.13431/j.cnki.immunol.j.20170044
GUERRERO FERNANDO, CIRAGAN ANNIKA, IWAÏ HIDEO: "Tandem SUMO fusion vectors for improving soluble protein expression and purification", PROTEIN EXPRESSION AND PURIFICATION, ACADEMIC PRESS, SAN DIEGO, CA., vol. 116, 1 December 2015 (2015-12-01), SAN DIEGO, CA. , pages 42 - 49, XP093006196, ISSN: 1046-5928, DOI: 10.1016/j.pep.2015.08.019
Attorney, Agent or Firm:
KANGXIN PARTNERS, P.C. (CN)
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