Title:
METHOD FOR OBTAINING RECOMBINANT BETA-1,4-GALACTOSIDASE BGAA FROM STREPTOCOCCUS PNEUMONIAE
Document Type and Number:
WIPO Patent Application WO/2020/197446
Kind Code:
A3
Abstract:
The invention relates to the field of biotechnology and addresses the problem of obtaining industrial quantities of highly purified enzymatically active recombinant β-1,4-galactosidase BgaA which is suitable for use in the remodeling stage to obtain therapeutically functional glycoprotein preparations with specified properties, for developing tools for high-sensitivity analysis of glycan chains of glycoproteins of plant, animal and microbial tissue. A method involves the use of metal chelate, pseudo-affinity, hydrophobic, anion-exchange, concentrating chromatography with subsequent polishing gel filtration, wherein a protein from the soluble fraction of cell lysate is purified using two stages of chromatography on the same Ni2+ metal chelate sorbent under different elution conditions and by pseudo-affinity chromatography in flow-through mode on Blue Sepharose. The use of the three stages mentioned above helps achieve maximum purification of the preparation for major contaminating impurities.
Inventors:
ERSHOV ALEKSANDR VIKTOROVICH (RU)
ERSHOVA OLGA ANATOLYEVNA (RU)
LEONOV VYACHESLAV SERGEEVICH (RU)
ERSHOVA OLGA ANATOLYEVNA (RU)
LEONOV VYACHESLAV SERGEEVICH (RU)
Application Number:
PCT/RU2020/050060
Publication Date:
November 19, 2020
Filing Date:
March 25, 2020
Export Citation:
Assignee:
LLC INTERNATIONAL BIOTECHNOLOGY CENTER GENERIUM (RU)
International Classes:
A61K38/47; C12N9/38
Domestic Patent References:
| WO1994012628A1 | 1994-06-09 |
Foreign References:
| US8236520B2 | 2012-08-07 |
Other References:
SUSANA CAMPUZANO H: "Cloning, Expression, and Characterization of a Peculiar Choline-Binding betaP-Galactosidase from Streptococcus mitis", APPL ENVIRON MICROBIOL., vol. 75, no. 18, September 2009 (2009-09-01), pages 5972 - 5980, XP055761012, Retrieved from the Internet [retrieved on 20200409]
DOROTHEA ZAHNER ET AL.: "The Streptococcus pneumoniae Beta-Galactosidase Is a Surface Protein", J. BACTERIOL, vol. 182, no. 20, 9 April 2020 (2020-04-09), pages 5919 - 5921, XP002529681, Retrieved from the Internet DOI: 10.1128/JB.182.20.5919-5921.2000
RYABTSEVA S. A. ET AL: "Sovremennye metody polucheniia Vgalaktozidaz. Nauka. Innovatsii. Tekhnologii,", 2014, pages 197 - 204, Retrieved from the Internet [retrieved on 20200409]
DOROTHEA ZAHNER ET AL.: "The Streptococcus pneumoniae Beta-Galactosidase Is a Surface Protein", J. BACTERIOL, vol. 182, no. 20, 9 April 2020 (2020-04-09), pages 5919 - 5921, XP002529681, Retrieved from the Internet
RYABTSEVA S. A. ET AL: "Sovremennye metody polucheniia Vgalaktozidaz. Nauka. Innovatsii. Tekhnologii,", 2014, pages 197 - 204, Retrieved from the Internet
Attorney, Agent or Firm:
PATENT & LAW FIRM 'YUS', LIMITED LIABILITY COMPANY (RU)
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