Large-scale preparation method of BCG-CpG-DNA

A technology of BCG and lysate, which is applied in the field of CpG-DNA and its preparation, can solve potential safety hazards and increase quality control testing items, etc.

Active Publication Date: 2014-03-19
NAT INST FOR FOOD & DRUG CONTROL
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Problems solved by technology

If it is used in production, not only will there be potential safety hazards, but there will be re

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  • Large-scale preparation method of BCG-CpG-DNA
  • Large-scale preparation method of BCG-CpG-DNA
  • Large-scale preparation method of BCG-CpG-DNA

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Embodiment Construction

[0048] The innovation and application significance of the present invention are described in detail below through specific examples and test results to help readers better understand the spirit and essence of the present invention, but it does not constitute a limitation to the implementation scope of the present invention.

[0049] Preparation of BCG-CpG-DNA

[0050] Step 1, bacteria culture:

[0051] The BCG strains were inoculated on potato Sutong medium. After culturing at 37°C for 15 days, transfer to improved liquid Sutong medium, and culture at 37°C for 14-20 days;

[0052] Wherein, the preparation method of potato Sutong culture medium can be:

[0053] Take and wash fresh potatoes (1), pierce them into cylinders with a piercer, and cut them into 4 cm long slopes with a knife;

[0054] Rinse the potato slopes with running drinking water for 1 hour;

[0055] Rinse the potato slant pieces with purified water;

[0056] Rinse the slant block with Sutong medium;

[005...

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Abstract

The invention provides a method for preparing Bacilli Calmette-Gurin CpG-DNA (BCG-CpG-DNA) by utilizing Q Sepharose HP ion exchange. The method comprises the following steps: carrying out culture breeding of BCG, cracking to obtain a BCG lysate, centrifuging the lysate, directly removing impurities comprising polysaccharides, proteins, RNA and the like from the centrifuged BCG lysate to obtain purified BCG-CpG-DNA. There is no organic solvent residual in the BCG-CpG-DNA prepared through the method.

Description

technical field [0001] The invention relates to CpG-DNA extracted from BCG (abbreviated as BCG-CpG-DNA) and a preparation method thereof. The BCG-CpG-DNA can be used as an adjuvant for vaccines for treatment and prevention. Background technique [0002] Adjuvants are indispensable components in vaccines. In terms of immune adjuvants, although the application of aluminum adjuvants has a history of 80 years and its safety has been tested for a long time, it has no obvious effect on cellular immunity, especially Vaccines based on cellular immunity, such as highly purified second-generation recombinant vaccines and third-generation DNA vaccines, have little effect, so people have been working on the research of new adjuvants. [0003] In recent years, CpG-DNA has become a research hotspot of new immune adjuvants. In addition to artificially synthesized CpG oligonucleotides (CpG ODN), prokaryotic DNA is the main source of CpG-DNA. BCG-CpG-DNA is a bacterial DNA fragment extract...

Claims

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Application Information

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IPC IPC(8): C12N15/10
Inventor 王国治赵爱华寇丽杰
Owner NAT INST FOR FOOD & DRUG CONTROL
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