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Bots antibacterial peptide adjuvant and preparation method thereof, and vaccine formulation containing adjuvant and use thereof

The technology of peptide adjuvant and antibacterial peptide is applied in the field of preparing vaccine preparations and achieves the effects of convenient use, simple preparation process and good safety.

Active Publication Date: 2014-07-09
ZHEJIANG UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, so far, the study of antimicrobial peptides used as vaccine adjuvants has not been reported

Method used

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  • Bots antibacterial peptide adjuvant and preparation method thereof, and vaccine formulation containing adjuvant and use thereof
  • Bots antibacterial peptide adjuvant and preparation method thereof, and vaccine formulation containing adjuvant and use thereof
  • Bots antibacterial peptide adjuvant and preparation method thereof, and vaccine formulation containing adjuvant and use thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0040] Embodiment 1: Preparation of fly maggot antimicrobial peptide

[0041] Take 1 kg of fly maggots, homogenize them in a high-speed tissue masher with 3% acetic acid aqueous solution for 5 min, place them in a refrigerator at 4°C overnight, centrifuge, and collect the supernatant; the precipitate is treated with 3% acetic acid aqueous solution in the same way. The two supernatants were combined, heated at 100 °C for 1 h, kept in a refrigerator at 4 °C, and filtered under reduced pressure. The filtrate was concentrated under reduced pressure and freeze-dried to obtain the fly maggot extract. Put the fly maggot extract on D101 macroporous adsorption resin chromatography column, elute with water and 55% ethanol in sequence, collect the 55% ethanol eluate, recover ethanol under reduced pressure, concentrate, and freeze-dry to obtain the fly maggot antimicrobial peptide. The fly maggot antimicrobial peptide was light yellow white powder; the ninhydrin reaction was positive; th...

Embodiment 2

[0042] Example 2 Hepatitis B Vaccine Preparation Containing Myiagia Antimicrobial Peptide Adjuvant

[0043] Weighed 500 μg of fly maggot antimicrobial peptide and 100 μg of recombinant hepatitis B virus surface antigen (rHBsAg), dissolved in 10 ml of normal saline, filtered through a 0.22 μm microporous membrane, and aseptically dispensed, 1 ml each. Each ml contains 50 μg of antimicrobial peptides and 10 μg of rHBsAg.

Embodiment 3

[0044] Example 3 Hepatitis B Vaccine Preparation Containing Aluminum Glue and Myiagia Antimicrobial Peptide Adjuvant

[0045] Weigh an appropriate amount of fly maggot antimicrobial peptide, dissolve it with physiological saline to make a solution with a concentration of 100 μg / ml, and filter it with a 0.22 μm microporous membrane. The antimicrobial peptide solution was mixed with an equal volume of aluminum-containing adjuvanted hepatitis B surface antigen vaccine stock solution (each ml containing 20 μg of rHBsAg and 500 μg of aluminum hydroxide), mixed evenly, and aseptically dispensed, 1 ml per tube. Each ml contains 50 μg of maggot antimicrobial peptide, 10 μg of rHBsAg and 250 μg of aluminum hydroxide.

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PUM

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Abstract

The invention relates to a bots antibacterial peptide adjuvant with an immune adjuvant role and a preparation method thereof, and a vaccine formulation containing the antibacterial peptide adjuvant and use thereof in preparation of the vaccine formulation. The antibacterial peptide is a polypeptide component extracted and separated from fly maggots, the cellular immunity and humoral immune response of immunized mice can be facilitated, the organism is induced to generate balanced Th1 type and Th2 type immunoreactions simultaneously, and the adjuvant characteristics better than those of the known aluminum plastic adjuvant in the prior art are displayed. The bots antibacterial peptide adjuvant can be used as an immunologic adjuvant for a plurality of vaccines to play an ideal immune effect. The vaccine of taking the antibacterial peptide as the adjuvant is simple in preparation process, simple and convenient in method, easy to control quality, convenient to use, and good in safety, and can be frozen to store.

Description

technical field [0001] The invention relates to a fly maggot antimicrobial peptide with immune adjuvant function extracted and separated from fly maggots, a preparation method thereof, a vaccine preparation containing the antibacterial peptide adjuvant, and their application in preparing vaccine preparations. Background technique [0002] Vaccination is the most economical and effective means to prevent and control infectious diseases. New vaccines such as DNA vaccines, recombinant vaccines, and synthetic peptide vaccines are a type of biological preparations that have developed rapidly in recent years. Compared with conventional vaccines, they have the advantages of high antigen purity, strong specificity, and good safety. However, these novel vaccines have weak immunogenicity and low immune potency [Oyston P, Robinso K. The current challenges for vaccine development. J Med Microbiol 2012; 61(7): 889–894]. At the same time, current vaccine research is facing important chal...

Claims

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Application Information

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IPC IPC(8): A61K39/39A61P31/00A61P37/04C07K1/14C07K1/16
Inventor 孙红祥
Owner ZHEJIANG UNIV
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