Synthetic peptide vaccines for foot-and-mouth disease

A free and sequential technology, applied in the field of synthetic peptide vaccines for foot-and-mouth disease, which can solve the problems of discontinuous high affinity and difficult identification

Inactive Publication Date: 2009-01-28
SHANGHAI SHEN LIAN BIOMEDICAL CORP
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

This research method is effective for positioning linear determinants, but the immune domain epitopes required for vaccine development are usually discontinuous epitopes with high affinity, which are difficult to identify by PEPSCAN method (Meloen et al., Ann Biol Clin, 1991, 49: 231- 242)

Method used

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  • Synthetic peptide vaccines for foot-and-mouth disease
  • Synthetic peptide vaccines for foot-and-mouth disease
  • Synthetic peptide vaccines for foot-and-mouth disease

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0149] Typical procedure for evaluating target antigenic peptides

[0150] FMDV involved in synthetic peptides

[0151] By Merrifield solid-phase synthesis technology, on the Applied Biosystems automatic matching peptide synthesizer (models 430, 431 and 433A), Fmoc compounds were used to synthesize peptides with sequences derived from the G-H loop region of FMDV VP1. can be provided for incorporation into a Selected amino acid mixtures at specific variable positions to prepare peptide constructs containing constructed synthetic antigen libraries (SSAL) for B or T cell epitopes, e.g., "O SSAL" (SEQ ID NO: 19) or "1, 4 , 9PALINDROMIC Th" (SEQ ID NO: 26). After assembly of the desired peptide immunogen, the resin is treated with trifluoroacetic acid following standard procedures to cleave the peptide from the resin and deblock the functional groups on the amino acid side chains. For cyclic peptides, the cleaved peptides were dissolved in 15% DMSO in water for 48 hours to promo...

Embodiment 2

[0163] Optimization of FMDV target antigen peptides by sequence information and immunostimulatory elements

[0164] Seven FMDV VP1 serotype A peptides are described in Table 7. Their corresponding VP1 amino acid sequences are aligned and numbered as in Table 1. The Th-carrying peptides of this group have an autologous FMDV Th identified as VP1 21-40 (SEQ ID NO: 24) described in Table 5, whose sequence is as follows: Glu-Thr-Gln-Ile-Gln-Arg-Arg- Gln-His-Thr-Asp-Val-Ser-Phe-Ile-Met-Asp-Arg-Phe-Val (SEQ ID NO: 24). The sequences of five of the seven constructs in Table 7 are detailed, represented as SEQ ID NOS: 1, 2, and 27-29. These peptides were synthesized and the resulting immune sera were evaluated for immunogenicity. Sera obtained 5 weeks after the initial immunization were analyzed for reactivity to the FMDV VP1 peptide by peptide-ELISA and by standard FMDV neutralization assays using strain A FP The ability to neutralize FMDV serotype A was analyzed as the target viru...

Embodiment 3

[0174] Application of hypervariable region consensus ordering of VP1 proteins that provide broad FMDV neutralization

[0175] synthetic constructs

[0176] Two synthetic circular constructs utilizing the consensus VP1 target antigenic site were designed following the consensus serotype O sequence shown in Table 1 (SEQ ID NO: 11). A consensus peptide was synthesized using an immunostimulatory element comprising the Inv domain (SEQ ID NO: 22) and the artificial SSAL Th (SEQ ID NO: 26) shown in Table 6. Other constructs were synthesized without additional immunostimulatory element sequences. The overall structures of these FMDV VP1 constructs are described as p2569a (SEQ ID NO: 11) in Table 8a, and p2570c in Table 8b (SEQ ID NO: 30).

[0177] The sera obtained after the initial immunization of hyperimmune guinea pigs for 3 and 5 weeks were evaluated by peptide ELISA for the reactivity of FMDV VP1 peptides, and by standard FMDV neutralization analysis, using FMDV strains AFP, O-...

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Abstract

The present invention relates to the use of a peptide composition as an immunogen, with each peptide contained therein comprising a target antigenic site derived from the VP1 capsid protein of Foot-and-Mouth Disease Virus (FMDV). The antigenic site is covalently linked to a helper T cell epitope and, preferably, to other immunostimulatory sequences, preferably by conventional peptide bond(s) through direct synthesis, for the prevention of FMDV infection and eradication of Food-and-Mouth Disease (FMD). More particularly, the present invention relates to the use of such peptide composition as an immunogen to elicit the production in animals including swine, cattle, sheep, goets and susceptible wild species, of high titer polyclonal antibodies that can effectively neutralize, in vitro, multiple strains or serotypes of FMDV, and to the use of such composition as a vaccine to prevent, and / or reduce the incidence of, FMDV infection regardless of serotype, and thus affect the eradication of FMD. The present invention also relates to the peptides used in the compositions, and to immunoassays and / or diagnostic kits containing one or more of these peptides, and methods of diagnosing FMDV inmammals using such materials.

Description

[0001] This application is the international application number "PCT / US99 / 13921", the international application date is June 21, 1999, the Chinese application date is December 15, 2000, the application number is "99807402.0", and the invention name is "for foot-and-mouth disease A divisional application for a synthetic peptide vaccine". field of invention [0002] The present invention relates to the use as an immunogen of a peptide composition comprising each peptide comprising a target antigenic site derived from the VPI capsid protein of foot-and-mouth disease virus (FMDV) covalently linked to a helper T Cellular epitopes, and preferably linked to other immunostimulatory sequences via direct synthesis, preferably via conventional peptide bonds, are useful for the prevention of FMDV infection and the eradication of Foot and Mouth Disease (FMD). [0003] More specifically, the present invention relates to the use of the peptide composition as an immunogen to induce high titer...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C07K14/09A61K39/00A61K39/12A61K39/245G01N33/53G01N37/00A61K39/125A61P31/12
CPCC12N2770/32122C12N2770/32134A61K39/00C07K14/005C12N15/52A61P31/12A61P31/14A61K39/245
Inventor 王长怡沈明
Owner SHANGHAI SHEN LIAN BIOMEDICAL CORP
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