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Neutralizing epitope from varicella-zoster virus (VZV) gE protein and antibody aiming the same

A protein and antibody technology, applied in the field of molecular virology and immunology, which can solve the problems of insufficient specificity of polyclonal antibodies and unsatisfactory therapeutic effect.

Active Publication Date: 2016-06-15
XIAMEN UNIV +1
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, the specificity of polyclonal antibody is not as good as that of monoclonal antibody, and its therapeutic effect is not satisfactory

Method used

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  • Neutralizing epitope from varicella-zoster virus (VZV) gE protein and antibody aiming the same
  • Neutralizing epitope from varicella-zoster virus (VZV) gE protein and antibody aiming the same
  • Neutralizing epitope from varicella-zoster virus (VZV) gE protein and antibody aiming the same

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specific Embodiment approach

[0170] The invention will now be described with reference to the following examples, which are intended to illustrate the invention, but not to limit it.

[0171] Unless otherwise specified, the molecular biology experiment methods and immunoassay methods used in the present invention are basically with reference to J.Sambrook et al., Molecular Cloning: A Laboratory Manual, 2nd Edition, Cold Spring Harbor Laboratory Press, 1989, and F.M.Ausubel et al., Refinement Molecular Biology Experimental Guide, 3rd Edition, John Wiley & Sons, Inc., 1995 The method described in; the use of restriction endonucleases was in accordance with the conditions recommended by the product manufacturer. The reagents whose sources are not indicated in the examples are all conventional reagents in the art or commercially available reagents. Those skilled in the art understand that the examples describe the present invention by way of example and are not intended to limit the scope of the claimed inven...

Embodiment 1

[0172] Embodiment 1: Preparation of recombinant VZV protein rgE

[0173] The invention uses an insect-baculovirus expression system to express and purify the extracellular segment of VZVgE glycoprotein. Specific steps are as follows:

[0174] 1. Construction of recombinant baculovirus

[0175] The present invention uses the p-Oka virus genome as a template, and uses the primers shown in Table 2 to perform PCR, thereby obtaining the gene encoding the gE glycoprotein extracellular segment (1-538aa). The amplified PCR product and pFastBac (pFB) vector were subjected to double enzyme digestion with BamHI and HindIII (both purchased from Japan TAKARA Company), respectively, and then the digested PCR product and vector were recovered and ligated. The ligation product was used to transform DH5α competent strains, and then the clones were selected for PCR, enzyme digestion and sequencing identification, and a positive clone containing the correct sequence was obtained, which was n...

Embodiment 2

[0183] Embodiment 2: Preparation of anti-VZVgE protein monoclonal antibody

[0184] mouse immunization

[0185] 1. Preparation of immunogen: The recombinant protein rgE (rgE) antigen was obtained by referring to the preparation method of the recombinant protein in Example 1. The 6-week-old BALB / c female mice were immunized with the purified rgE protein, and 5 female mice (numbered: mouse 1 to mouse 5) were immunized with an immunization dose of 100 μg / mouse. The antigen used for primary immunization was mixed with Freund's complete adjuvant (purchased from Sigma). After the initial immunization, booster immunization was performed once every two weeks, and the antigen used in the booster immunization was mixed with Freund's incomplete adjuvant (purchased from Sigma), and the booster immunization was performed three times in total. On day 11 after the third booster immunization, blood was collected. The last booster immunization was carried out 72 hours before the fusion, a...

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Abstract

The invention relates to a neutralizing epitope peptide (or a variant thereof) from varicella-zoster virus virus (VZV) gE protein, a recombinant protein containing the neutralizing epitope peptide (or the variant thereof) and a carrier protein, and applications of the neutralizing epitope peptide (or a variant thereof) and the recombinant protein, and also relates to an antibody aiming to the neutralizing epitope peptide, a cell strain producing the antibody, and the applications thereof, and also relates to a vaccine containing the neutralizing epitope peptide and the recombinant protein, a medicine composition including the antibody, and the applications thereof, e.g., the application for preventing and / or treating VZV infection or one or more diseases and symptoms related to the infection.

Description

technical field [0001] The present invention relates to the fields of molecular virology and immunology. Specifically, the present invention relates to neutralizing epitope peptides (or variants thereof) from varicella-zoster virus gE protein, recombinant proteins comprising such neutralizing epitope peptides (or variants thereof) and carrier proteins, As well as the use of such neutralizing epitope peptides (or variants thereof) and recombinant proteins. The present invention also relates to antibodies against such neutralizing epitope peptides, cell lines producing the antibodies, and their uses. The present invention also relates to vaccines comprising said neutralizing epitope peptides or recombinant proteins, pharmaceutical compositions comprising said antibodies, and their uses, for example, they can be used to prevent and / or treat varicella-zoster virus infection or One or more diseases or symptoms associated with the infection. Background technique [0002] Varice...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C07K7/08C07K14/04C07K19/00C12N7/04C12N15/38C07K16/08C12N5/20C12N15/70C12N15/81C12N15/83C12N15/86C12N5/10A61K39/395A61P31/22G01N33/68G01N33/577
CPCA61K39/395C07K7/08C07K14/005C07K16/088C07K19/00C12N2710/16023C12N2710/16034G01N33/577G01N33/68
Inventor 程通刘剑朱瑞王玮叶祥忠李益民夏宁邵
Owner XIAMEN UNIV
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