Conjugate of influenza A virus conservative peptides M2e and virus-like particles, and application thereof

A type A influenza virus, virus-like technology, applied in the direction of antiviral agents, medical preparations with non-active ingredients, medical preparations containing active ingredients, etc., can solve problems such as no good protective effect and dependence on chicken embryos , to achieve the effect of easy large-scale production and short preparation time

Inactive Publication Date: 2013-05-08
苏州科贝生物技术有限公司
View PDF4 Cites 5 Cited by
  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

The current influenza vaccines mainly target the surface glycoproteins hemagglutinin (HA) and neuraminidase (Neuraminidase, NA) that induce neutralizing antibodies, but there are two factors that restrict the effectiveness of seasonal influenza vaccines in preventing and controlling influenza pandemics. Application in the epidemic: one is whether the vaccine strain matches

Method used

the structure of the environmentally friendly knitted fabric provided by the present invention; figure 2 Flow chart of the yarn wrapping machine for environmentally friendly knitted fabrics and storage devices; image 3 Is the parameter map of the yarn covering machine
View more

Image

Smart Image Click on the blue labels to locate them in the text.
Viewing Examples
Smart Image
  • Conjugate of influenza A virus conservative peptides M2e and virus-like particles, and application thereof
  • Conjugate of influenza A virus conservative peptides M2e and virus-like particles, and application thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0018] Example 1. Preparation of M2e-Q β-VLP conjugate and reference substance M2e-keyhole limpet hemocyanin (KLH) conjugate

[0019] Step 1. Preparation of bacteriophage virus-like particle Q β-VLP: inoculate the strain Escherichia coli DH5α / pETQ β-CP into LB medium and cultivate until the appropriate bacterial solution reaches the OD value, add the inducer IPTG to induce for several hours, and collect the bacteria Cells were disrupted by ultrasound, and after lysis, the supernatant and pellet were collected by centrifugation. The formed multimer was purified using Sepharose CL-4B chromatography column.

[0020] Step 2, the synthesis of M2e polypeptide: adopt the solid-phase synthesis method to utilize the polypeptide synthesizer to automatically synthesize the conserved sequence M2e of human influenza A virus M2 protein

[0021] Step 3. The peptide segment M2e is coupled to the bacteriophage virus-like particle Q β-VLP and keyhole limpet hemocyanin (KLH) respectively.

[0...

Embodiment 2

[0023] Embodiment two, ELISA method detection mouse serum antibody titer

[0024] Twenty 6-week-old C57BL / 6 female mice were randomly divided into 4 groups, with an average of 5 mice in each group. The groups were divided into M2e polypeptide group, M2e-QβVLP group, M2e-KLH group and control group. See Table 1 below for the immunization methods, doses, time intervals, and times.

[0025] Table 1: Immunization strategy

[0026] time

times

dosage

Immunization method

0 weeks

1st

50ug

subcutaneous more

2 weeks

2nd

50ug

subcutaneous more

4 weeks

the 3rd time

50ug

subcutaneous more

[0027] 100ul of mouse serum was collected before each immunization. Store at -20°C.

[0028] Dissolve M2e polypeptide, VLP, and KLH in 0.1mM carbonate buffer solution (pH 9.6), make 20ug / mL antigen coating solution, coat 96-well plates with 100ul per well, and coat overnight at 4°C. quilt. Then it was blocke...

Embodiment 3

[0030] Embodiment 3, mouse protective experiment

[0031] 56 8-week-old C57BL / 6 female mice were randomly divided into 4 groups, with an average of 14 in each group. The groups were divided into placebo group, M2e-Q βVLP group, M2e-KLH group, and M2e group. See Table 2 below for the immunization methods, doses, time intervals, and times.

[0032] Table 2: Immunization schedule for C57BL / 6 mice

[0033] time

[0034] On the 28th day, 7 of the mice in each group were inoculated with 4 times the semi-lethal dose of influenza A strain (A / PR / 8 / 34[PR8, H1N1] PR8 strain, and the other 7 mice were inoculated with 4 times the semi-lethal dose of Influenza A virus NIB-64A / Perth / 16 / 2009[H3N2] strain, followed by closely observing the survival status of the mice every day, continued to observe for 28 days. The number of survival mice was counted. The experimental results are shown in Table 3. The experimental results show that, The M2e-QβVLP group was immunized twice with 50...

the structure of the environmentally friendly knitted fabric provided by the present invention; figure 2 Flow chart of the yarn wrapping machine for environmentally friendly knitted fabrics and storage devices; image 3 Is the parameter map of the yarn covering machine
Login to view more

PUM

No PUM Login to view more

Abstract

The invention relates to a conjugate of virus-like particles (VLP) of oligopeptide vector Q[beta] bacteriophage with target immunotherapy and conservative peptides M2e of human influenza virus M2 protein. The conjugate is mainly used for preventing human influenza A. The conjugate is coupled by a Sulfo-SMCC method which comprises a first step of preparing the virus-like particles (VLP) of the Q[beta] bacteriophage; a second step of preparing the virus-like particles (VLP) activated by Sulfo-SMCC; and a third step of preparing the conjugate of M2e-VLP. The conjugate can produce high titer protective antibody in mice and cross-protect the attack of various subtypes of the influenza A viruses.

Description

technical field [0001] The invention belongs to the field of biotechnology, and specifically relates to a conjugate formed by immunologically coupling an immune targeting therapy carrier with a conserved short peptide M2e of influenza A virus. The conjugate can cross-protect the challenge of various subtypes of influenza A virus. Background technique [0002] The preparation of a universal influenza vaccine is a hot spot in current influenza vaccine research. Influenza is an acute respiratory infectious disease that seriously endangers human health. Influenza vaccination is the most effective means of protection. The current influenza vaccines mainly target the surface glycoproteins hemagglutinin (HA) and neuraminidase (Neuraminidase, NA) that induce neutralizing antibodies, but there are two factors that restrict the effectiveness of seasonal influenza vaccines in preventing and controlling influenza pandemics. Application in the epidemic: one is whether the vaccine strai...

Claims

the structure of the environmentally friendly knitted fabric provided by the present invention; figure 2 Flow chart of the yarn wrapping machine for environmentally friendly knitted fabrics and storage devices; image 3 Is the parameter map of the yarn covering machine
Login to view more

Application Information

Patent Timeline
no application Login to view more
IPC IPC(8): A61K39/385A61K39/145A61K47/48A61P31/16A61K47/64
Inventor 姬云昇文国刘红海许峰马严堵芸倩
Owner 苏州科贝生物技术有限公司
Who we serve
  • R&D Engineer
  • R&D Manager
  • IP Professional
Why Eureka
  • Industry Leading Data Capabilities
  • Powerful AI technology
  • Patent DNA Extraction
Social media
Try Eureka
PatSnap group products

Browse by: Latest US Patents, China's latest patents, Technical Efficacy Thesaurus, Application Domain, Technology Topic.

© 2024 PatSnap. All rights reserved.Legal|Privacy policy|Modern Slavery Act Transparency Statement|Sitemap