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Recombinant antigen protein with function of easily activating B cells and preparation method thereof

A technology for recombining antigens and B cells, applied in the field of immunology, can solve problems such as uneven assembly, increased activation of B cells, epitope blocking, etc., and achieve the effects of reducing the vaccination dose, prolonging the duration of antibodies, and increasing the molecular weight of proteins

Active Publication Date: 2021-04-20
龙湖现代免疫实验室
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

This strategy can theoretically increase the probability of activating B cells, but in fact, this strategy does not increase the probability of activating B cells, but instead leads to problems with the uniformity and stability of VLPs vaccines, such as epitope blocking and uneven assembly etc. Therefore, the research and development of safe and efficient new subunit vaccines is very important

Method used

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  • Recombinant antigen protein with function of easily activating B cells and preparation method thereof
  • Recombinant antigen protein with function of easily activating B cells and preparation method thereof
  • Recombinant antigen protein with function of easily activating B cells and preparation method thereof

Examples

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Effect test

Embodiment 1

[0141] Embodiment 1 classical swine fever virus recombinant antigen

[0142] This example provides a CSFV recombinant antigen that can easily activate B cells and a preparation method thereof.

[0143] 1. Determine the target protein:

[0144] Classical swine fever (CSF) is a highly contagious, acute and fatal infectious disease caused by classical swine fever virus (CSFV). The CSFV genome is a single-stranded positive-strand RNA, which is a spherical enveloped virus. The structure of CSFV was analyzed by literature search and bioinformatics prediction ( Figure 4 ) found that the four major structural proteins of CSFV are C, Erns (gp44 / 48), E1 (gp33), and E2 (gp55). Among them, the molecular weight of structural protein E2 is about 55kDa, which is a glycosylated envelope protein, mainly located on the surface of virus particles, and plays an important role in the process of virus invasion into cells; at the same time, E2 protein is also the main protective antigen of CSFV,...

Embodiment 2

[0178] Example 2 Newcastle disease virus recombinant antigen

[0179] This example provides a recombinant Newcastle disease virus antigen that can easily activate B cells and a preparation method thereof.

[0180] 1. Determine the target protein:

[0181] Newcastle disease virus (NDV) is an approximately spherical particle with an envelope, and often has an irregular shape due to the damage of the envelope, and filaments of different lengths with a cross-sectional diameter of about 100 nm can be seen. Through literature search and bioinformatics prediction analysis of NDV virus structure ( Figure 15). The surface of the viral envelope is covered with 8nm-long filaments, and the inside of the virion is a coiled nucleocapsid with a diameter of about 17nm. The six specific structural proteins of NDV are L protein (large protein, large protein), NP (nucleuo protein, nucleoprotein), P protein (phosphor protein, phosphoprotein), M protein (matrix protein, matrix protein), F prot...

Embodiment 3

[0222] Example 3 Novel Coronavirus Recombinant Antigen

[0223] This example provides a novel coronavirus recombinant antigen that is easy to activate B cells and a preparation method thereof.

[0224] 1. Determine the target protein:

[0225] Analysis of SARS-CoV-2 virus structure through literature search and bioinformatics prediction ( Figure 28 ), the four main structural proteins of SARS-CoV-2 are spike protein (Spike protein, S), envelope protein (Envelop protein, E), matrix protein (Membrane protein, M) and nucleocapsid protein (Nucleocapsid protein , N). Among them, the S protein is an important class of glycoproteins on the surface of coronaviruses, including key epitopes of coronaviruses, and is the main target for pathogen detection and vaccine research. Therefore, S protein was chosen as the target protein.

[0226] 2. Structural analysis of target protein and identification of key epitopes

[0227] 1) Target protein secondary structure prediction

[0228] T...

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Abstract

The invention relates to a recombinant antigen protein with a function of easily activating B cells and a preparation method thereof. According to the recombinant antigen protein provided by the invention, two or more antigen proteins containing at least one same epitope peptide are linked through a linker peptide to prepare a dimer or polymer recombinant antigen, and the length of the linker peptide enables the distance between the at least two same epitopes to be 3-50 nm. According to the recombinant antigen, at least two of the same epitopes in the antigen protein can be recognized by BCR on B cells at the same time, so that the BCR is cross-linked, and the B cells are more effectively activated. The invention further provides a preparation method of the recombinant protein antigen. The method is universal, can be used for preparing the recombinant antigen with remarkably improved B cell activation efficiency, and can be widely applied to the fields of vaccine antigen preparation, antibody preparation and the like.

Description

technical field [0001] The invention belongs to the technical field of immunology, and in particular relates to a recombinant antigenic protein capable of activating B cells and a preparation method thereof. Background technique [0002] For more than a century, vaccines have made an important contribution to the control of infectious diseases. Over the years, vaccine research has mostly relied on unlimited passages to obtain weakened strains, which is uncertain and blind. With the development of protein structure and the emergence of protein structure analysis techniques such as cryo-electron microscopy, people have mastered the structural information of many pathogen surface proteins. In addition, with the development of basic immunology, the immunogenic design of antigens is becoming more and more reasonable. Structure-based antigen design greatly reduces uncertainty in vaccine development. [0003] At present, many recombinant products such as subunit vaccines and VLP...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C07K14/18C07K14/165C07K14/125C12N15/82
Inventor 张改平王爱萍丁培阳刘红亮陈玉梅
Owner 龙湖现代免疫实验室
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