Common Specific Epitope of Boca Virus (HBoV), Its Application and Antibody

A Boca virus-specific technology, applied in the field of Boca virus, can solve the problem that there is no relevant report on the epitope research of Boca virus VP2 protein

Active Publication Date: 2017-11-10
INST OF PATHOGEN BIOLOGY CHINESE ACADEMY OF MEDICAL SCI
View PDF2 Cites 0 Cited by
  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, there is no relevant report on the epitope research of Boca virus VP2 protein

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
  • Common Specific Epitope of Boca Virus (HBoV), Its Application and Antibody
  • Common Specific Epitope of Boca Virus (HBoV), Its Application and Antibody
  • Common Specific Epitope of Boca Virus (HBoV), Its Application and Antibody

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0026] Example 1. Identification of the immunodominant epitope of Boca virus VP2 protein

[0027]The HBoV1-3VP2 gene used herein was derived from bocavirus positive diarrhea stool samples 111-BJ07, 211-BJ07 and 46-BJ07 (Genbank numbers: JQ240469, JQ240470 and HM132056, respectively). Random phage libraries of HBoV1-3VP2 genes were constructed according to the method disclosed in the literature, and named as libraries HBoV1-VP2, HBoV2-VP2 and HBoV3-VP2 respectively. Then use the mouse anti-HBoV1-3VP2 antibody prepared by the applicant and Boca virus positive human serum to carry out affinity screening to the random phage library of HBoV1-3VP2 gene (random phage library preparation method and screening method refer to Khurana S, Suguitan AL Jr, Rivera Y , et al. Antigenic fingerprinting of H5N1avian influenza using convalescent sera and monoclonal antibodies reveals potential vaccine and diagnostic targets. PLoS Med, 20096: e1000049). figure 1 Shown are the epitope distribution...

Embodiment 2

[0028] Example 2. Immunogenicity and antigenic characteristics of immunodominant epitopes

[0029] 2.1 Coupling of peptides to KLH

[0030] According to the distribution of Boca virus antigenic epitopes and the sequence analysis of HBoV1VP2 protein, two dominant antigenic epitope peptides, P1 and P2 (sequences are shown in SEQ ID NO: 1 and SEQ ID NO: 2, respectively), were synthesized and coupled with KLH respectively. Combined to increase the immunogenicity of the polypeptide (Table 1), it was used to immunize mice. The synthesis of the peptide and the coupling with KLH were entrusted to Shanghai Bioengineering Technology Co., Ltd.

[0031] Table 1. Sequences of the peptide-KLH conjugates used for immunization

[0032] name

Location

sequence

P1-KLH

1-20 a

KLH-CMSDTDIQDQQPDTVDAPQNT

P2-KLH

162-180 a

KLH-CEHAYPNASHPWDEDVMPDL

[0033] a Indicates the corresponding position of the polypeptide in the VP2 protein (sequence show...

Embodiment 3

[0041] Example 3. Fine mapping of P1 and P2 synthetic peptide epitopes

[0042] In order to further determine the key amino acid residues in the P1 and P2 peptides, we synthesized a series of peptides, the results are shown in Table 2 and Table 3.

[0043] Table 2 Peptides used for fine mapping of P1 peptides

[0044]

[0045] Table 3 Peptides used for fine mapping of P2 peptides

[0046]

[0047] Competitive ELISA detection: Dilute P1 and P2 with coating solution (carbonate buffer at pH 9.6: 1.59g sodium carbonate, 2.93g sodium bicarbonate, diluted with water to 1000mL) and then coated A microtiter plate (product of Costar Company) was placed at 4° C. overnight, adding blocking solution (1% BSA / PBS) and incubated at 37° C. for 2 hours, washed 5 times with 0.1% PBST buffer, and dried the washing solution. Simultaneously different concentrations (3 0 ×10pmol / well, 3 1 ×10pmol / well, 3 2 ×10pmol / well, 3 3 ×10pmol / well, 3 4 ×10pmol / well, 3 5 ×10pmol / well, 3 6 ×10pmo...

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

PropertyMeasurementUnit
Sensitivityaaaaaaaaaa
Sensitivityaaaaaaaaaa
Login to view more

Abstract

The invention discloses a boca virus (HBoV) common specific epitope peptide, its application and its antibody. Specifically, the specific epitope peptide is MSDTDIQDQQPDTVDAPQNT or EHAYPNASHPWDEDVMPDL.

Description

technical field [0001] The present invention relates to boca virus, especially epitopes that can be used for the diagnosis of diseases caused by the virus, its application and its antibody. Background technique [0002] Human bocavirus (huamn bocavirus, HBoV) is a newly discovered virus in recent years, and it is the second parvovirus known to infect humans. Four genotypes (HBoV1-4) of HBoV have been found, and HBoV1 was found in a nasopharyngeal sample of a child with acute respiratory infection in Sweden in 2005. Subsequently, three more bocaviruses (HBoV2-4) were successively found in stool samples from 2009 to 2010. [0003] Human bocavirus infection is distributed worldwide, and the virus can be detected not only in respiratory tract and stool samples, but also in serum, tonsil, saliva and urine. The analysis of PCR detection results of respiratory samples shows that the detection rate of HBoV1 is about 2-19% depending on the country, region, detection method and popu...

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
Patent Type & Authority Patents(China)
IPC IPC(8): C07K7/08C07K16/08G01N33/68
Inventor 王健伟郭丽王雅英G·弗奈特
Owner INST OF PATHOGEN BIOLOGY CHINESE ACADEMY OF MEDICAL SCI
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