US-20260124296-A1 - NINE-COMPONENT ANTIGEN AFRICAN SWINE FEVER SUBUNIT VACCINE

US20260124296A1US 20260124296 A1US20260124296 A1US 20260124296A1US-20260124296-A1

Abstract

The present disclosure belongs to the field of biotechnology, and specifically relates to a nine-component antigen African swine fever subunit vaccine. The present disclosure first provides an African swine fever virus antigen protein combination composed of the African swine fever virus P34 protein, P30 protein, P54 protein, A104R protein, E165R protein, C129R protein, P72 protein, X protein, and Y protein. This African swine fever virus antigen protein combination can induce a strong immune response in the host. Furthermore, the present disclosure provides a nine-component antigen African swine fever subunit vaccine including the aforementioned African swine fever virus antigen protein combination. The nine-component antigen African swine fever subunit vaccine exhibits good immunoprotection rates against challenge with the parental virulent African swine fever virus strain, poses no biosafety risks, overcomes the difficulty that existing African swine fever subunit vaccines domestically and internationally cannot provide effective immunoprotection for pigs.

Inventors

Haixue ZHENG
Wei Zhang
Yajun Li
Yang Yang
Zhengwang SHI
Jijun He
Ye Jin
Dan Li
Zixiang ZHU
Ruoqing MAO
Rongzeng HAO
Yi Ru
Guicai Zhang
Wen Dang
Xusheng Ma
Xiaodong Qin
Xiangtao Liu
Huanan LIU
Jianhong GUO
Hong Tian
Weijun Cao
Bingzhou LU
Fan Yang
Chaochao SHEN

Assignees

LANZHOU VETERINARY RESEARCH INSTITUTE, CHINESE ACADEMY OF AGRICULTURAL SCIENCES

Dates

Publication Date: 20260507
Application Date: 20251009
Priority Date: 20230410

Claims (15)

1 . An African swine fever virus (ASFV) antigen protein combination, wherein the ASFV antigen protein combination consists of a P34 protein, a P30 protein, a P54 protein, an A104R protein, an E165R protein, an X protein, a C129R protein, a P72 protein, and a Y protein: wherein the X protein is a fusion protein of a DP96R protein and a P12 protein; and the Y protein is a fusion protein of an extracellular domain of a P22 protein and an extracellular domain of a P17 protein; the P34 protein, the P30 protein, the P54 protein, the A104R protein, the E165R protein, the X protein, the C129R protein, the P72 protein, and the Y protein are in a ratio of (1-6):(1-6):(1-6):(1-6):1:(1-6):(1-6):1:(1-6); the concentration of each of the P34 protein, the P30 protein, the P54 protein, the A104R protein, the E165R protein, the X protein, the C129R protein, the P72 protein, and the Y protein is greater than or equal to 90 μg/mL; the amino acid sequence of the P34 protein is set forth in SEQ ID NO: 1; the amino acid sequence of the P30 protein is set forth in SEQ ID NO: 3; the amino acid sequence of the P54 protein is set forth in SEQ ID NO: 5 or SEQ ID NO: 7; the amino acid sequence of the A104R protein is set forth in SEQ ID NO: 9; the amino acid sequence of the E165R protein is set forth in SEQ ID NO: 11; the amino acid sequence of the DP96R protein is set forth in SEQ ID NO: 13; the amino acid sequence of the fusion protein of the DP96R protein and the P12 protein is set forth in SEQ ID NO: 15; the amino acid sequence of the C129R protein is set forth in SEQ ID NO: 17; the amino acid sequence of the P72 protein is set forth in SEQ ID NO: 19; the amino acid sequence of the P17 protein is set forth in SEQ ID NO: 21; the amino acid sequence of the P22 protein is set forth in SEQ ID NO: 23; the amino acid sequence of a fusion protein of a fragment of the P17 protein and a fragment of the P22 protein is set forth in SEQ ID NO: 25; and the amino acid sequence of the fusion protein of the extracellular domain of the P22 protein and the extracellular domain of the P17 protein is set forth in SEQ ID NO: 27.
2 . The ASFV antigen protein combination according to claim 1 , wherein the ASFV is a genotype II ASFV.
3 . The ASFV antigen protein combination according to claim 2 , wherein the genotype II ASFV is ASFV CN/GS 2018.
4 . The ASFV antigen protein combination according to claim 1 , wherein the P34 protein, the P30 protein, the P54 protein, the A104R protein, the E165R protein, the X protein, the C129R protein, the P72 protein, and the Y protein are in a ratio of (1-2):1:(1-2):1:1:(1-2):1:1:1.
5 . The ASFV antigen protein combination according to claim 4 , wherein the P34 protein, the P30 protein, the P54 protein, the A104R protein, the E165R protein, the X protein, the C129R protein, the P72 protein, and the Y protein are in a ratio of 1:1:(1-2):1:1:(1-2):1:1:1.
6 . The ASFV antigen protein combination according to claim 1 , wherein the concentration of 2010.2 each of the P34 protein, the P30 protein, the P54 protein, the A104R protein, the E165R protein, the X protein, the C129R protein, the P72 protein, and the Y protein is greater than or equal to 150 μg/mL.
7 . The ASFV antigen protein combination according to claim 6 , wherein the concentration of each of the P34 protein, the P30 protein, the P54 protein, the A104R protein, the E165R protein, the X protein, the C129R protein, the P72 protein, and the Y protein is from 150 μg/mL to 2400 μg/mL.
8 . The ASFV antigen protein combination according to claim 7 , wherein the concentrations of the P34 protein, the P30 protein, the P54 protein, the A104R protein, the E165R protein, the X protein, the C129R protein, the P72 protein, and the Y protein are 200 μg/mL, 200 g/mL, 400 μg/mL, 200 μg/mL, 200 μg/mL, 400 μg/mL, 200 μg/mL, 200 μg/mL, and 200 μg/mL, respectively.
9 . The ASFV antigen protein combination according to claim 7 , wherein the concentrations of the P34 protein, the P30 protein, the P54 protein, the A104R protein, the E165R protein, the X protein, the C129R protein, the P72 protein, and the Y protein are 200 μg/mL, 200 μg/mL, 200 μg/mL, 200 μg/mL, 200 μg/mL, 400 μg/mL, 200 μg/mL, 200 μg/mL, and 200 μg/mL, respectively.
10 . The ASFV antigen protein combination according to claim 7 , wherein the concentrations of the P34 protein, the P30 protein, the P54 protein, the A104R protein, the E165R protein, the X protein, the C129R protein, the P72 protein, and the Y protein are 400 μg/mL, 200 μg/mL, 200 μg/mL, 200 μg/mL, 200 μg/mL, 400 μg/mL, 200 μg/mL, 200 μg/mL, and 200 μg/mL, respectively.
11 . The ASFV antigen protein combination according to claim 1 , wherein the P34 protein, the P30 protein, the P54 protein, the A104R protein, the E165R protein, the X protein, and the C129R protein are obtained by expression in an Escherichia coli system; the P72 protein is obtained by expression in an insect cell expression system; and the Y protein is obtained by expression in a Chinese hamster ovary (CHO) expression system.
12 . A use of the ASFV antigen protein combination according to any one of claims 1-11 in the manufacture of a medicament for preventing or treating an ASFV infection.
13 . A use of the ASFV antigen protein combination according to any one of claims 1-11 in the manufacture of a biological product for preventing an ASFV infection.
14 . A nine-component antigen African swine fever subunit vaccine, wherein the nine-component antigen African swine fever subunit vaccine consists of the ASFV antigen protein combination according to any one of claims 1-11 and a pharmaceutically acceptable adjuvant.
15 . The nine-component antigen African swine fever subunit vaccine according to claim 14 , wherein the adjuvant comprises one or more selected from the group consisting of: a chemical immunological adjuvant, a microbial immunological adjuvant, a plant-based immunological adjuvant, and a biochemical immunological adjuvant.

Description

CROSS-REFERENCE TO RELATED APPLICATIONS This application is a continuation of International Application No. PCT/CN2024/086656, filed on Apr. 8, 2024, which claims priority to Chinese Patent Application No. 202310374606.0, filed on Apr. 10, 2023. All of the aforementioned applications are incorporated herein by reference in their entireties. SEQUENCE LISTING This application contains Sequence Listing which has been submitted in XML format via EFS-Web and is hereby incorporated by reference in its entirety. Said XML copy is named Nine component antigen African swine fever subunit vaccine.xml, created on Sep. 25, 2025, and has a size of 37,620 bytes. TECHNICAL FIELD The present disclosure belongs to the field of biotechnology, and specifically relates to a nine-component antigen African swine fever subunit vaccine. BACKGROUND African swine fever (ASF) is the “number one killer” threatening the global swine industry, with a prevalence history of over one hundred years. In August 2018, ASF was introduced into China, severely impacting the domestic swine industry and causing unprecedented losses. To date, no commercialized ASF vaccine has been made available, presenting a critical challenge and pain point for the swine industry and constituting a major national demand. Scientific research both domestically and internationally indicates that ASF inactivated vaccines, subunit vaccines, and live vector vaccines still fail to address the issue of immunogenic efficacy. Although gene-deleted vaccines resolve the immunogenic efficacy concern, they pose risks associated with residual virulence and biosafety. The African swine fever virus (ASFV) possesses a large genome encoding over 150 viral proteins, most of which have unclear functions. The antigens responsible for inducing and stimulating the immune system remain poorly understood, which represents a key obstacle in developing an African swine fever subunit vaccine with ideal immunogenic efficacy. Regarding subunit vaccines, Chinese Patent CN115814071A discloses a subunit vaccine comprising African swine fever virus p34, p14, C129R, DP96R, A104R, p54, p17, p22, P72, and p30 proteins; Chinese Patent CN115702928A discloses a subunit vaccine comprising African swine fever virus p34, p14, C129R, DP96R, A104R, p54, p22, P72, and p30 proteins; Chinese Patent CN112472801A discloses a subunit vaccine comprising African swine fever p30, p54, p72, and B602L proteins; Chinese Patent CN111658768A discloses a subunit vaccine comprising the African swine fever virus surface envelope protein CD2V and at least one protein selected from the group consisting of African swine fever virus P72 protein, African swine fever virus P30 protein, and African swine fever virus P54 membrane structural protein. Although the aforementioned studies have disclosed subunit vaccines with different antigen compositions, none have systematically evaluated the immunoprotective efficacy of the vaccines. Further research by the applicant's team has also revealed that certain antigen components involved in the aforementioned subunit vaccines may exhibit antibody-dependent enhancement (ADE) effects, which could severely compromise the protective efficacy of the subunit vaccines against virulent strains of African swine fever. The term “ADE effect” refers to a process wherein virus-specific antibodies bind to the virus, and the virus-antibody complexes subsequently bind via the Fc segment to cells expressing Fc receptors (FcR) on their surface, thereby facilitating viral entry into these cells and enhancing the infectivity of the virus. Based on the above issues, the applicant provides a nine-component antigen African swine fever subunit vaccine. The subunit vaccine demonstrates ideal protective efficacy against challenge with a parental virulent strain of African swine fever. SUMMARY In response to the above technical problems, the present disclosure, through extensive experimental screening and evaluation research, has unexpectedly discovered a nine-component antigen African swine fever subunit vaccine. The subunit vaccine not only induces a superior immune response in the organism and poses no biosafety risks, but also provides ideal protective efficacy upon challenge with a parental virulent strain of African swine fever. Specifically, the present disclosure includes the following content. In a first aspect, the present disclosure provides an ASFV antigen protein combination, wherein the ASFV antigen protein combination consists of a P34 protein, a P30 protein, a P54 protein, an A104R protein, an E165R protein, an X protein, a C129R protein, a P72 protein, and a Y protein; wherein the X protein is a DP96R protein or a fusion protein of the DP96R protein and a P12 protein; and the Y protein is a P22 protein, or a P17 protein, or a combination of the P22 protein and the P17 protein, or a fusion protein of the P22 protein and the P17 protein, or a fusion protein of a fragment of the P22 protein and a fragmen