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Purification method of lentivirus

A purification method, lentivirus technology, applied in the biological field, can solve the problem of low recovery rate

Active Publication Date: 2020-07-24
GUANGDONG FAPON BIOPHARMA INC
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Although the existing lentivirus production and preparation process can meet the requirements of large-scale, high purity, and high titer, there is still the problem of low recovery rate

Method used

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  • Purification method of lentivirus
  • Purification method of lentivirus
  • Purification method of lentivirus

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Experimental program
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specific Embodiment

[0060] The following will be described in detail in conjunction with specific embodiments. The following examples, unless otherwise specified, do not include other components except unavoidable impurities. The drugs and instruments used in the examples are all routine choices in the art unless otherwise specified. The experimental methods for which specific conditions are not indicated in the examples are implemented according to conventional conditions, such as the conditions described in literature, books or the method recommended by the manufacturer.

[0061] The materials in the following examples are commercially available products, wherein the nuclease is a product of Qihai Forte Biotechnology, the sorbitol is a product of Adamas Reagent with a purity of >99%, the maltose is a product of Sinopharm Shanghai Test with a purity of >99%, and manna Alcohol is a product of Adamas Reagent with a purity of >99%, sucrose is a product of Shanghai Sangong with a purity of >99%, an...

Embodiment 17～ Embodiment 23

[0076] (1) At 4°C, collect the supernatant of the lentiviral culture medium, centrifuge at 3000g for 5 minutes, collect the centrifuged supernatant to obtain the centrifugate, then mix it according to the volume ratio of nuclease and centrifugate 1:5000, and then use a 0.45μm pore size filter Membrane filtration to obtain primary purified liquid.

[0077] (2) Concentrate the primary purified solution obtained in step (1) with an ultrafiltration membrane bag with a pore size of 100 kD at 4° C. to obtain an ultrafiltrate.

[0078] (3) After mixing the ultrafiltrate in step (2) with the additive, let it stand for 30 minutes to obtain a pre-displacement product. The components of the additives and the concentrations of the additives in the pre-displacement are shown in Table 2.

[0079] Table 2

[0080]

[0081]

[0082] (4) The ultrafiltrate was washed with PBS buffer solution containing corresponding additives, and the volume ratio of the ultrafiltrate and buffer solutio...

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Abstract

The invention relates to a slow virus purification method. According to the slow virus purification method, at least one of additives selected from saccharose, sorbitol, maltose, trehalose and mannitol is used in the ultrafiltration concentration process of purification for assisting concentration. With the adoption of the slow virus purification method, recovery rate in the slow virus purification process can be increased.

Description

technical field [0001] The invention relates to the field of biotechnology, in particular to a method for purifying lentivirus. Background technique [0002] Lentivirus has a very wide application in the fields of gene therapy and cell therapy, and is an important exogenous gene delivery carrier. Gene therapy and cell therapy have extremely high requirements on the purity and titer of lentivirus. Although the existing lentivirus production and preparation process can meet the requirements of large-scale, high purity, and high titer, there is still the problem of low recovery rate. Contents of the invention [0003] Based on this, it is necessary to provide a purification method to improve the recovery rate of lentivirus. [0004] A method for purifying lentivirus, using at least one additive selected from sucrose, sorbitol, maltose, trehalose and mannitol to assist concentration during the ultrafiltration concentration process of purification. [0005] It has been confi...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C12N7/02C12R1/93
CPCC12N7/00C12N2740/15051
Inventor 芦迪刘云鹏王先进张宏涛
Owner GUANGDONG FAPON BIOPHARMA INC