Title:
PURIFICATION METHOD FOR AND APPLICATION OF GMP-GRADE RETROVIRAL VECTOR
Document Type and Number:
WIPO Patent Application WO/2022/262206
Kind Code:
A1
Abstract:
Disclosed is a method for purifying a retrovirus (retroviral vector), comprising the following steps: A), micro-filtering a cell culture supernatant containing a retrovirus so as to remove cell debris from the cell culture supernatant, and obtaining a micro-filtered virus solution; B), treating the microfiltered virus solution with a nuclease so as to degrade host DNA residues into small DNA fragments, and obtaining an enzyme-digested virus solution; C), ultra-filtering the enzyme-digested virus solution such that the retrovirus is retained in a retentate, collecting the retentate, and obtaining an ultra-filtered virus solution; and D), performing low-speed centrifugation treatment on the ultra-filtered virus solution so as to precipitate the retrovirus, collecting the precipitate, and obtaining a virus that has not undergone sterilization. The retrovirus product obtained by the described method ensures the purity of the retrovirus solution, and at the same time, the titer of the concentrated virus solution can reach 107IP/ml, which meets downstream processing requirements.
Inventors:
ZHU JIANGAO (CN)
LI JIAJIA (CN)
YANG WENJUN (CN)
LI JIAJIA (CN)
YANG WENJUN (CN)
Application Number:
PCT/CN2021/134470
Publication Date:
December 22, 2022
Filing Date:
November 30, 2021
Export Citation:
Assignee:
CARBIOGENE THERAPEUTICS CO LTD (CN)
International Classes:
C12N7/02; C12N15/867; C12R1/93
Foreign References:
| CN113373120A | 2021-09-10 | |||
| CN111876393A | 2020-11-03 | |||
| US20210009966A1 | 2021-01-14 | |||
| CN111876392A | 2020-11-03 | |||
| CN105980571A | 2016-09-28 |
Other References:
LOO ET AL.: "Production of High Titer cGMP-Grade SIN Gamma-Retroviral Vectors by Transfection in a Closed System Bioreactor", BLOOD, vol. 112, no. 11, 16 November 2008 (2008-11-16), XP086679747, ISSN: 0006-4971, DOI: 10.1182/blood.V112.11.3539.3539
ZENG RONG, LI JIN, ZHOU SHENG, ET AL : "Purification Packaging, Titer and Detection of C-myc Retrovirus Expression Vector", PROGRESS OF ANATOMICAL SCIENCES, vol. 6, no. 3, 20 September 2000 (2000-09-20), XP093015202, ISSN: 1006-2947, DOI: 10.16695/j.cnki.1006-2947.2000.03.018
ZENG RONG, LI JIN, ZHOU SHENG, ET AL : "Purification Packaging, Titer and Detection of C-myc Retrovirus Expression Vector", PROGRESS OF ANATOMICAL SCIENCES, vol. 6, no. 3, 20 September 2000 (2000-09-20), XP093015202, ISSN: 1006-2947, DOI: 10.16695/j.cnki.1006-2947.2000.03.018
Attorney, Agent or Firm:
HANGZHOU YUYANG UNIONPATENT AGENCY CO., LTD. (CN)
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