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CpGODN adjuvant and application thereof in antibody production

An adjuvant and antibody technology, applied in recombinant DNA technology, resistance to vector-borne diseases, DNA/RNA fragments, etc., can solve the problems that the target protein cannot be fully recognized and the cost of monoclonal antibodies is high.

Pending Publication Date: 2022-08-02
安徽环球基因科技有限公司
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

In vitro diagnostic reagents, immunoturbidimetric reagents can use monoclonal antibodies or polyclonal antibodies, the cost of using monoclonal antibodies is high, because only limited epitopes can be recognized, and complete recognition of the target protein in the sample cannot be guaranteed

Method used

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  • CpGODN adjuvant and application thereof in antibody production
  • CpGODN adjuvant and application thereof in antibody production
  • CpGODN adjuvant and application thereof in antibody production

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0025] Application test of CpG ODN adjuvant in antibody production

[0026] Step 1. Antigen preparation:

[0027] Human procalcitonin (PCT) recombinant protein recombinantly expressed in Escherichia coli was used as the immunogen, and the antigen concentration was 1 mg / mL; the basic immune antigen was 500ul recombinant PCT protein, mixed with 500ul Freund's complete adjuvant (Sigma, F5881), and carried out 1 minute phacoemulsification, repeated 3 times; 500ul recombinant PCT protein was used as the booster immunization antigen, mixed with 500ul incomplete Freund's adjuvant (Sigma, F5506), 1 minute phacoemulsification, repeated 3 times;

[0028] Step 2. Preparation of CpG ODN mixed adjuvant:

[0029] Synthesis of CpG ODN-1, CpG ODN-2, CpG ODN-3, CpG ODN-4, CpG ODN-5, CpG ODN-6, CpGODN-7, CpG ODN-8, CpG ODN-9, CpG ODN-10. Use ddH2O to dissolve into 5mg / mL mother solution, mix 0.1mL of each, and mix by vortex; before use, mix CpG ODN and Lipofectamine 2000 (Thermo, 11668019) in...

Embodiment 2

[0046] Application test of CpG ODN adjuvant in antibody production

[0047] Compared with Example 1, the immunization protocol in step 3 was replaced with the following protocol, and other steps were the same:

[0048] Optimized cycle group: booster immunization was performed every other week after basic immunization, and blood was collected one week after the fourth immunization. The basic immunization was inoculated with Freund's complete adjuvant and antigen emulsified, and CpG ODN mixed adjuvant was injected alone 3 days later. The booster immunization was inoculated with incomplete Freund's adjuvant and antigen emulsified, and CpG ODN mixed adjuvant was injected alone at the same time.

Embodiment 3

[0056] Result analysis:

[0057] 1. Comparison of titers: Table 2 shows the titer data of the antibodies obtained in Example 1-2 and Comparative Example 1-2.

[0058] Table 2

[0059]

[0060] From the data in Table 2, it can be seen that the antibody titer results obtained in Example 1-2 are better than those obtained in Comparative Examples 1-2, and the antibody titer obtained in Example 2 is better than that obtained in Example 1. price result.

[0061] 2. Yield comparison: Table 3 shows the titer data of the antibodies obtained in Example 1-2 and Comparative Example 1-2.

[0062] table 3

[0063]

[0064] From the data in Table 3, it can be seen that the yield of antibody produced in Example 2 is higher than that of Example 1 and Comparative Examples 1-2.

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Abstract

The invention relates to a CpG ODN adjuvant and application thereof in antibody production, and belongs to the technical field of polyclonal antibody production. The application of the CpG ODN adjuvant in antibody production comprises the following steps: synthesizing according to one or more of sequences of CpGODN-1, CpG ODN-2, CpG ODN-3, CpG ODN-4, CpG ODN-5, CpG ODN-6, CpGODN-7, CpG ODN-8, CpG ODN-9 and CpG ODN-10, mixing, and mixing with a cationic liposome reagent in an equal volume, thereby obtaining the CpG ODN mixed adjuvant for antibody production. By using the CpG ODN mixed adjuvant provided by the invention, the conventional animal immune cycle can be shortened from 10 weeks to 4 weeks. The titer of the produced polyclonal antibody product is obviously higher than that of a control group, and the yield of the antibody is also higher than that of the control group.

Description

technical field [0001] The invention belongs to the technical field of polyclonal antibody production, and in particular relates to a CpG ODN adjuvant and its application in antibody production. Background technique [0002] Polyclonal antibody refers to immunizing animals with antigens containing multiple antigenic determinants, which can stimulate multiple B cell clones in the body to produce different antibodies against multiple antigenic epitopes. The obtained immune serum is actually a mixture of multiple antibodies. Among in vitro diagnostic reagents, immunoturbidimetric reagents can use monoclonal antibodies or polyclonal antibodies. The cost of using monoclonal antibodies is high because only limited epitopes can be recognized, and complete recognition of the target protein in the sample cannot be guaranteed. Therefore, most of the immunoturbidimetric reagents on the market currently use polyclonal antibodies. Polyclonal antibodies have low cost and can be mass-prod...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N15/11C07K16/06
CPCC12N15/11C07K16/06Y02A50/30
Inventor 缪连军刘宗文潘红张磊张志鹏刘倩朱桃花
Owner 安徽环球基因科技有限公司
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