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Application of natural antibacterial peptide QHA in preparation of immunologic adjuvant

A natural antibacterial and immune adjuvant technology, applied in the direction of antibodies, antineoplastic drugs, antibody medical components, etc., can solve the problems of different antimicrobial peptide sequences and immunobiological effects that cannot be generalized

Active Publication Date: 2020-07-17
SUZHOU UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, different antimicrobial peptides have different sequences, and their principles of regulating the host's immune response are also different, and the immunobiological effects of different antimicrobial peptides as adjuvants cannot be generalized.

Method used

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  • Application of natural antibacterial peptide QHA in preparation of immunologic adjuvant
  • Application of natural antibacterial peptide QHA in preparation of immunologic adjuvant
  • Application of natural antibacterial peptide QHA in preparation of immunologic adjuvant

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0029] The preparation of embodiment 1QHA

[0030] (1) Synthesize the full sequence (shown in SEQ ID No.1) derived from the reptiles Qinghuanhai snake QHA with an automatic peptide synthesizer (433A, Applied Biosystems), and desalt and purify it by HPLC reverse-phase column chromatography.

[0031] (2) Molecular weight was determined by matrix-assisted laser desorption ionization time-of-flight mass spectrometry (MALDI-TOF).

[0032] (3) The purity of the purified QHA was identified by high performance liquid chromatography (HPLC), the molecular weight was determined by matrix-assisted laser desorption ionization time-of-flight mass spectrometry (MALDI-TOF), the isoelectric point was determined by isoelectric focusing electrophoresis, and the amino acid was determined by an automatic amino acid sequencer sequence structure.

[0033] The amino acid sequence of the natural antimicrobial peptide QHA of the present invention is shown in SEQ ID No.1. It consists of 30 amino acids...

Embodiment 2

[0034] Example 2QHA is used as an immune adjuvant to detect the impact of mouse antibody production

[0035] The experiments were divided into antigen group, antigen / QHA group and blank group (injected with the same volume of PBS), with 3 mice in each group.

[0036] The SARS-CoV-2 new coronavirus S1 protein antigen (provided by Suzhou Sige Bio) and the antimicrobial peptide QHA were dissolved in PBS to 1 mg / ml and 0.5 mg / ml, respectively, to obtain the antigen solution and QHA solution. In the antigen group, each mouse was immunized with 100 μl of antigen solution each time, and the immunization method was intraperitoneal immunization.

[0037] In addition, the above-prepared antigen solution was mixed with the QHA solution at a volume ratio of 1:1, and the resulting mixture was placed on a shaking table at 37°C and incubated for 35 minutes with shaking at 100 rpm / min to obtain an antigen / QHA mixture. In the antigen / QHA group, each mouse was immunized with 200 μl of antigen / ...

Embodiment 3

[0041] Embodiment 3 QHA combined with Freund's adjuvant as immune adjuvant for the detection of the impact of mouse antibody production

[0042] The experiments were divided into antigen / Freund's adjuvant group, antigen / QHA group and antigen / Freund's adjuvant / QHA group, with 3 mice in each group.

[0043] The SARS-CoV-2 new coronavirus S1 protein antigen (provided by Suzhou Sige Biological) and the antibacterial peptide QHA were dissolved in PBS to 1mg / ml and 0.5mg / ml to obtain the antigen solution and QHA solution. Then the antigen solution and Freund's adjuvant (volume ratio 1:1), antigen solution and QHA solution (volume ratio 1:1), antigen solution, Freund's adjuvant and QHA solution (volume ratio 1:1:1) were mixed , to obtain antigen / Freund's adjuvant mixed solution, antigen / QHA mixed solution and antigen / Freund's adjuvant / QHA mixed solution. When mixing, place the mixture on a vortex shaker for 35 minutes to vigorously shake to fully emulsify the Freund's adjuvant.

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Abstract

The invention relates to an application of natural antibacterial peptide QHA in preparation of an immunologic adjuvant. The immunologic adjuvant is used for improving the immune titer of an antibody.In addition, the QHA and a Freund adjuvant have a synergistic effect when being combined for use. The invention discloses the novel application of the natural antibacterial peptide QHA. The natural antibacterial peptide QHA can be used for preparing the immunologic adjuvant for improving the immune titer of the antibody. The QHA as immunologic adjuvant for improving the immune titer of the antibody has wide application prospects in the fields of preparation of antibodies, vaccines, immunotherapy drugs, tumor vaccines or immune activators and the like.

Description

technical field [0001] The invention relates to an immune adjuvant, in particular to the application of a natural antibacterial peptide QHA in preparing the immune adjuvant. Background technique [0002] Immunological adjuvants are referred to as adjuvants for short, that is, non-specific immune proliferators, which refer to auxiliary substances that can enhance the body's immune response to antigens or change the type of immune response together with antigens or pre-injected into the body. Adjuvants may or may not be immunogenic. The immunobiological effects of adjuvants include enhancing immunogenicity, enhancing antibody titers, changing the type of antibody production, causing or enhancing delayed hypersensitivity reactions, etc. Because immune adjuvants can enhance the immune response, they are widely used. Its uses mainly include: ①As a non-specific immune enhancer, it is used for adjuvant treatment of anti-infection and anti-tumor; ②It can enhance specific immune re...

Claims

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Application Information

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IPC IPC(8): A61K39/39A61K38/16A61K38/17A61K39/395A61P35/00A61P37/04
CPCA61K39/39A61K38/16A61K38/17A61K39/39558A61P35/00A61P37/04A61K2039/55516A61K2300/00
Inventor 王义鹏欧阳建红章铭辉张登登陈燕
Owner SUZHOU UNIV
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