Construction and application of fusion protein vaccine platform

Abstract

To provide vaccine platforms that enhance biological response to viral, bacterial, or tumor antigens.SOLUTION: The present invention provides a vaccine comprising a fusion protein containing an interferon-target antigen-immunoglobulin Fc region (or antibody) and a Th cell helper epitope. The present invention also relates to the use of a fusion protein containing an interferon-target antigen-immunoglobulin Fc region (or antibody) and a Th cell helper epitope for preparing a preventive or therapeutic composition. The vaccine of the present invention is produced by a eukaryotic cell expression system, and wild-type and various mutant antigen vaccines can be prepared, and a strong immune response can be induced in the living body by inoculation via an immune route such as subcutaneous / muscle or nasal cavity. The vaccine of the present invention can be used as a preventive or therapeutic vaccine.SELECTED DRAWING: None

Description

[Technical field]

[0001] The present invention belongs to the field of genetic engineering and biomedical technology, and specifically relates to a vaccine, for example a vaccine comprising a fusion protein containing interferon-target antigen-immunoglobulin Fc region (antibody) as a backbone. The vaccine of the present invention is used as a vaccine platform to prevent Hepatitis B virus (HBV) infection, HPV, EBV, HIV, SARA-COV2, influenza virus infection, and prevent the development of HPV, EBV-related tumors, as well as to treat chronic hepatitis B (CHB) infection, and treat HBV, HP, EBV-related tumors. [Background technology]

[0002] Globally, approximately 257 million people are chronically infected with the virus, and approximately 88,700 people die each year from HBV-related end-stage liver disease, including liver failure, cirrhosis, and hepatocellular carcinoma. [1-3] Approximately 30% of cases of cirrhosis and 40% of cases of hepatocellular carcinoma (HCC) are due to HBV. Hepatitis B virus infection remains a major public health problem worldwide. [4] However, there is still no effective treatment for chronic hepatitis B. Existing HBV treatments mainly include antiviral drugs (nucleoside / nucleotide analogs) and interferon. Although these have certain therapeutic effects, they usually cannot induce an effective immune response and therefore cannot completely eliminate HBV infection. And the side effects of long-term medication are relatively large, and antiviral drugs can also develop drug resistance. Chronic HBV infection is one of the major diseases threatening human health, and it is urgent to explore effective immunotherapy strategies for chronic hepatitis B, and the development of a therapeutic vaccine for chronic hepatitis B is of great social and economic importance.

[0003] Seasonal influenza causes severe illness in 1 to 4 million people each year and 200,000 to 500,000 deaths. [5]Vaccines are the best way to prevent and control influenza. Vaccines can reduce the incidence of disease and the severity of infection, especially in young children and the elderly, who are at risk of influenza complications. Although currently approved influenza vaccines provide adequate protection in healthy young adults, there are still some issues that need to be addressed. For example, the production pathways for some vaccines, such as inactivated and attenuated influenza vaccines, rely on chicken embryos. A drawback of these vaccines is that if the circulating strain is of avian origin, a disease outbreak can increase the demand for both vaccines and chicken embryos, creating problems with the supply of chicken embryos. [6] Another drawback is that these vaccines take a lot of time to produce. Older people are more susceptible to severe influenza virus syndromes. At the same time, standard vaccines are generally less effective in older people, whose immune systems gradually weaken with age. [7] In light of the problems with current influenza vaccines, there is an urgent need to develop an influenza vaccine that is highly immunogenic, does not depend on chicken embryos, and can be rapidly produced in response to influenza virus epidemics.

[0004] Severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) is the pathogen that caused the coronavirus disease 2019 (COVID) pandemic. Clinical symptoms caused by SARS-CoV-2 are primarily asymptomatic infection, mild flu-like symptoms, pneumonia and severe acute respiratory distress syndrome, which in severe cases can lead to death in infected patients. [8] Currently, there is no specific medicine for the new coronavirus, and a vaccine is the fundamental measure to control and end the COVID-19 pandemic. [9] Additionally, the emergence of new variants of the coronavirus poses new challenges to existing candidate vaccines and epidemic containment.

[10] Therefore, in the current pandemic situation, a vaccine that is potent and can act against novel coronavirus variants is urgently needed.

[0005] When an antigen is linked to the Fc region of an immunoglobulin, its half-life is significantly increased and the Fc region of the immunoglobulin can bind to Fc receptors on the surface of antigen-presenting cells, facilitating antigen processing and presentation by the antigen-presenting cells. [11-13] Type I interferons are antiviral cytokines that have many biological activities, one of which is the stimulation of immune cells.

[14] IFNα can potently induce the differentiation and activation of human DC cells.

[15] After type I interferon acts on immature DCs, it promotes the expression of MHC molecules and costimulatory molecules on the cell surface of DCs, such as MHC class I, CD80, and CD86, thereby enhancing the ability of DCs to activate T cells. [16-18] It has been reported that type I interferons can enhance the antigen-presenting capacity of DCs after infection with vaccinia virus and lymphocytic chorioencephalitis virus (LCMV). [19-21] In addition, type I interferon can promote the migration of DCs to lymph nodes by upregulating the expression of chemokine receptors after acting on DCs, thereby promoting the activation of T cells. [22、23] Recently, more and more studies have shown that type I interferons can be used as immune adjuvants. A study by Le Bon et al. showed that type I interferons exhibited a strong immune adjuvant effect in mice when immunized with a weak immunogen, inducing long-lasting antibodies and immune memory.

[24] We also found that DC cells are the main cell population on which type I interferon acts, and antibodies can be used to target and deliver vaccines to DCs, stimulating their activation and cross-presentation function, and further enhancing the activity and efficacy of vaccines.

[0006] There is a need in the present invention to provide a vaccine platform that enhances the body's response to viral, bacterial, or tumor antigens. Summary of the Invention

[0007] Vaccines are an effective way to prevent and control infectious disease outbreaks, and there are various kinds of vaccines, one of which is the protein subunit vaccine. In general, simple protein subunit vaccines often have limited use due to their generally low immunogenicity. Therefore, a universal protein subunit vaccine platform is urgently needed. Based on the influence of immunoglobulin Fc region and type I interferon on the immune system, the present inventors have specifically proposed an interferon α-viral antigen, bacteria or tumor-immunoglobulin Fc region fusion protein vaccine platform to enhance the body's response to virus, bacteria or tumor antigens. The present invention provides a type I interferon-protein antigen-immunoglobulin Fc vaccine platform, in which type I interferon acts on antigen-presenting cells to make them mature and migrate, so as to better play the role of antigen presentation and T cell activation, while the Fc portion of the vaccine platform can bind to the Fc receptor on the surface of antigen-presenting cells to promote the uptake of antigens by antigen-presenting cells, and further aid the function of antigen-presenting cells. The present inventors proposed the fusion of Th cell helper epitopes, which can further improve the immune response effect of type I interferon-protein antigen-immunoglobulin Fc vaccines and are an important component of the vaccine. The present inventors proposed that antibodies such as anti-PD-L1 can be used to replace Fc to target and deliver vaccines to DCs, stimulate DC activation and cross-presentation function, and further enhance the activity and efficacy of the vaccine. As a novel vaccine platform, the present invention can be used as a preventive and therapeutic vaccine for diseases such as viral infections, bacterial infections, or tumors.

[0008] In some embodiments, the present invention provides a vaccine comprising a fusion protein (Th epitope-added) containing an interferon-target antigen-immunoglobulin Fc region (or antibody). In some embodiments, the present invention further provides the use of a fusion protein (Th epitope-added) containing an interferon-target antigen-immunoglobulin Fc region (or antibody) for preparing a prophylactic or therapeutic composition or kit (e.g., a drug or vaccine composition or kit). The vaccine of the present invention can be produced by a eukaryotic cell expression system and can be administered by an immunization route such as subcutaneous / muscle or nasal. In the fusion polypeptide of the present invention, the antibody (abbreviated as Ab) as a structural unit thereof is not particularly limited, and may include, for example, a complete antibody, an antibody fragment such as an antibody heavy chain or light chain, or a single chain antibody, or may be a DC-targeting activating antibody including antibodies such as anti-PD-L1, anti-DEC205, and anti-CD80 / 86.

[0009] In some embodiments, the target antigen described herein is not particularly limited and can be any suitable antigen. In some embodiments, the target antigen described herein can be, for example, a tumor antigen and / or a pathogen antigen (e.g., a viral or bacterial antigen). In some embodiments, the target antigen described herein can be, for example, a tumor antigen, for example, a tumor cell highly expressed protein molecule, for example, human epidermal growth factor receptor 2 (HER2 / neu), epidermal growth factor receptor (EGFR).

[0010] In some embodiments, the target antigen used in the vaccines provided herein may be, for example, a mutant target antigen that differs from the wild type. In some embodiments, the target antigen described herein may be, for example, a mutant of a tumor antigen and / or a pathogen antigen (e.g., a viral or bacterial antigen). In some embodiments, the target antigen may be, for example, the full-length or S1 region of the SARS-COV-2 virus S protein, e.g., the target antigen may be an antigen set forth in SEQ ID NO: 76 or SEQ ID NO: 77. As used herein, a wild-type target antigen refers to an immunogenic protein expressed by a virus or other infectious agent or tumor that is encoded by a wild-type gene (a wild-type gene refers to an allele that is predominant in nature and is often used as a standard control gene in biological experiments), e.g., the Spike protein (S protein) derived from the original wild-type strain of SARS-CoV-2. As used herein, a mutant target antigen (mutant) refers to a mutant viral protein expressed by a mutant viral strain encoded by a mutant gene mutated from a wild-type gene. For example, point mutations in different mutant novel coronavirus S proteins that have been discovered include NTD region 69-70 deletion, Y144 deletion, 242-244 deletion, L18F, D80A, D215, R246I mutations, RBD region K417, E484, N501Y and other mutations, L452R mutation, T478K mutation, D614G, H655Y mutations. For example, these point mutations are present in various combinations in mutated novel coronaviruses derived from the UK B.1.1.7 (Alpha) variant, the South African B.1.351 (Beta) variant, the Brazilian P1 (Gamma) variant, the Indian B.1.617, B.1.617.1 (Kappa), B.1.617.2 (Delta), and B.1.617.3 variants, and the California B.1.429 variant.In some embodiments, the mutant target antigen may include, for example, natural point mutations / deletion mutations / gain mutations / truncations, artificial point mutations / deletion mutations / gain mutations / truncations, any combination of natural or artificial mutations, subtypes generated after mutation, where the target antigen may be a tumor antigen, a pathogen antigen, such as a viral (e.g., SARS-COV-2) or bacterial antigen. In some embodiments, the target antigen used in the vaccine provided by the present invention is a mutant viral antigen, for example, the mutant viral antigen may be a mutant of SARS-COV-2, for example, a natural point mutation / deletion mutation / gain mutation / truncated form, an artificial point mutation / deletion mutation / gain mutation / truncated form, any combination of natural or artificial mutations, or a subtype generated after mutation, of a SARS-COV-2 protein (e.g., at least one selected from the group consisting of S protein, N protein, M protein, and E protein), for example, the mutant viral antigen may be a mutant of the full-length S protein, S1 region, or RBD region, for example, the mutant viral antigen may be a mutant of the NT of the S protein of SARS-COV-2. The mutant viral antigen may include at least one mutation selected from the group consisting of D region 69-70 deletion, Y144 deletion, 242-244 deletion, L18F, D80A, D215, R246I mutation, RBD region K417, E484, N501Y mutation, L452R mutation, T478K mutation, D614G, H655Y mutation. For example, the mutant viral antigen may include a mutation derived from the UK B.1.1.7 (Alpha) mutant strain, South African B.1.351 (Beta) mutant strain, Brazilian P1 (Gamma) mutant strain, Indian B.1.617, B.1.617.1 (Kappa), B.1.617.2 (Delta), B.1.617.3 mutant strain, or California B.1.429 mutant strain. For example, the mutant viral antigen may include a mutation derived from SEQ ID NO: For example, the mutant viral antigen may comprise a variant of a sequence shown in any one of SEQ ID NO:79, SEQ ID NO:80, SEQ ID NO:81, SEQ ID NO:82, SEQ ID NO:83, SEQ ID NO:84.As used herein, unless otherwise indicated or clearly limited by context, target antigens referred to herein generally include wild-type target antigens and mutant target antigens.

[0011] The present invention aims to provide a vaccine platform (Th epitope-added) consisting of interferon (IFN) and a tumor, bacterial or viral antigen (Hepatitis B virus Pres1 antigen, SARS-COV2 RBD antigen, influenza HA antigen, human papillomavirus HPV E7 antigen, Hepatitis B virus surface antigen (HBsAg) antigen or peptide, Varicella-Zoster virus (VZV) gE antigen, Epstein-Barr virus (EBV) EBNA1 / LMP2 / gp350, Herpes simplex virus 2 (HSV-2) gD antigen, AIDS virus (HIV) gp120 antigen) and an immunoglobulin Fc region (or antibody). The fusion protein may be a homodimeric protein or a heterodimeric protein. When the fusion protein is a dimer, the interferon, target antigen, and immunoglobulin Fc region (or antibody Ab) as structural units may be present in the first polypeptide chain and / or the second polypeptide chain. The form in which each structural unit exists is not particularly limited. For example, they may exist simultaneously in one chain, or any one or more structural units may exist in one chain and another one or more structural units may exist in another chain.

[0012] The interferon according to the invention can be selected from the group consisting of type I interferon, type II interferon, type III interferon, such as IFN-α, IFN-β, IFN-γ, IFN-λ1 (IL-29), IFN-λ2 (IL-28a), IFN-λ (IL-28b) and IFN-ω. The IFN can be of human or murine origin. The interferon according to the invention is preferably IFN-α (SEQ ID NO: 1, SEQ ID NO: 21, SEQ ID NO: 22) as type I interferon.

[0013] The immunoglobulin Fc region according to the present invention can be selected from the group consisting of the constant region amino acid sequences of IgG1, IgG2, IgG3, IgG4 and / or IgM, preferably IgG1 (SEQ ID NO: 2, SEQ ID NO: 23, SEQ ID NO: 24).

[0014] The fusion polypeptide of the present invention may also optionally include one or more Th cell helper epitopes and / or linking fragments (linkers). For example, when the fusion protein is a dimer, the fusion protein may also optionally include one or more Th cell helper epitopes and / or linking fragments in any one or two chains (i.e., the first polypeptide chain and / or the second polypeptide chain) of the homodimer or heterodimer. As known to those skilled in the art, each structural unit of the fusion protein can be linked by a suitable linking fragment (linker). The linking fragment that can be used in the vaccine of the present invention is not particularly limited and can be any suitable peptide fragment known in the art. The linking fragment of each structural unit described in the present invention may be a flexible polypeptide sequence, and may be linking fragments 1 and 2, for example, those shown in the amino acid sequences of SEQ ID NO: 4 and SEQ ID NO: 25.

[0015] The N-terminus of each polypeptide sequence consisting of each structural unit according to the present invention contains a corresponding signal peptide capable of promoting protein secretion, for example, as shown in the amino acid sequence of SEQ ID NO:5.

[0016] Preferred antigens according to the present invention include Hepatitis B Pres1 antigen, including the ad subtype (SEQ ID NO: 6), ay subtype (SEQ ID NO: 26), HBV HBsAg antigen (subtypes and peptides), including the adr subtype (SEQ ID NO: 7), adw subtype (SEQ ID NO: 27), ayw subtype (SEQ ID NO: 28), SARA-COV2 RBD antigen (SEQ ID NO: 8), Influenza virus HA antigen (SEQ ID NO: 9), HPV E7 antigen (SEQ ID NO: 10), Herpes virus VZV-gE antigen (SEQ ID NO: 91), EBV-gp350 antigen (SEQ ID NO: 92), HSV-2-gD antigen (SEQ ID NO: 93).

[0017] The homodimeric protein according to the present invention comprises a first polypeptide and a second polypeptide, and the first polypeptide and the second polypeptide are completely identical. The first polypeptide and the second polypeptide are, in the order from N-terminus to C-terminus, IFN-tumor or viral antigen (Hepatitis B Pres1 antigen, SARS-COV2 RBD antigen, Influenza HA antigen, HPV E7 antigen, HBsAg antigen, VZV-gE antigen, EBV EBNA1 / LMP2 / gp350, HSV-2-gD antigen, HIV gp120 antigen)-immunoglobulin Fc region, or are Pan epitope-containing polypeptides, and comprise the amino acid sequences shown in SEQ ID NO: 11, 12, 13, 14, 29, 30, 31, 32, 38, 39, 40, 47, 48, 49, 50, 51, 56, 57, 59, 58, 65, 66, 67, 68.

[0018] The heterodimer according to the present invention comprises a first polypeptide and a second polypeptide, wherein the first polypeptide and the second polypeptide are not the same polypeptide, and the first polypeptide is an IFN-immunoglobulin Fc region, respectively, from the C-terminus to the N-terminus, and comprises the amino acid sequence shown in SEQ ID NO: 15, 33, 42, 51, 60, 69, and the second polypeptide is a tumor or viral antigen (Hepatitis B Pres1 antigen, SARS-COV2 RBD antigen, Influenza HA antigen, HPV E7 antigen, VZV-gE antigen, EBV EBNA1 / LMP2 / gp350, HSV-2-gD antigen, HIV gp120 antigen)-immunoglobulin Fc region, respectively, from the C-terminus to the N-terminus, and comprises the amino acid sequence shown in SEQ ID NO: Includes the amino acid sequences shown in 16, 17, 18, 19, 34, 35, 36, 37, 43, 44, 45, 46, 52, 53, 54, 55, 61, 62, 63, 64, 70, 71, 72, and 73.

[0019] The present invention also provides amino acid sequences encoding the above-mentioned IFN-tumor or viral antigen (Hepatitis B Pres1 antigen, HBsAg antigen or peptide, SARS-COV2 RBD antigen, influenza HA antigen, HPV E7 antigen, VZV-gE antigen, EBV EBNA1 / LMP2 / gp350, HSV-2-gD antigen, HIV gp120 antigen)-immunoglobulin Fc vaccine platform.

[0020] The present invention also relates to nucleotide segments encoding said vaccine platforms and fusion proteins.

[0021] The present invention also relates to a method for preparing the fusion protein or vaccine platform, for example comprising the steps of: (1) constructing an expression vector comprising a coding gene encoding the fusion protein or vaccine platform, preferably, the expression vector is a pEE12.4 expression vector; (2) constructing a host cell comprising the expression vector by transiently transfecting a host cell, preferably the host cell is a 293F cell; (3) culturing the host cells and harvesting the cell supernatant; (4) purifying the fusion protein or vaccine platform by Protein A / G affinity chromatography column purifying protein;

[0022] The present invention also includes applications of the vaccine platform, wherein the vaccine platform can be used as a hepatitis B prophylactic vaccine; the vaccine platform can be used as a hepatitis B therapeutic vaccine; the vaccine platform can be used as an influenza prophylactic vaccine; the vaccine platform can be used as a SARA-COV2, influenza, HPV, VZV, EBV, HSV-2, HIV prophylactic vaccine; and the vaccine platform can be used as an HPV, EBV-associated tumor prophylactic vaccine.

[0023] The present invention includes adjuvants used by the vaccine platform, including aluminum adjuvant (Alum), Toll-like receptor 4 activator ligand MPLA, Toll-like receptor 9 ligand, M59, oligodeoxynucleotides (CpG-ODN), and Freund's adjuvant.

[0024] The present invention includes the clinical use of said vaccine platform in combination with the Hepatitis B virus envelope protein HBsAg vaccine as an HBV therapeutic vaccine in the treatment of chronic Hepatitis B virus infection.

[0025] The present invention includes the clinical use of the combination of said vaccine platform with a nucleoside or nucleotide analogue as an HBV therapeutic vaccine in the treatment of chronic Hepatitis B virus infection.

[0026] The invention includes the use of said vaccine platform in combination with antiviral drugs and other therapies, such as for HBV, influenza, SARA-COV2, HPV, VZV, EBV, HSV-2, HIV prophylactic or therapeutic vaccines, and includes the use of said vaccine platform in combination with antiviral and tumor drugs and therapies, such as for HBV, HPV, EBV associated tumor prophylactic or therapeutic vaccines.

[0027] The present invention includes multivalent combination vaccines consisting of the vaccine platform as one component of a vaccine with another virus or pathogen or tumor vaccine.

[0028] The present invention includes any fusion protein vaccine of said vaccine platform immunized in a sequential or simultaneous immunization procedure with an adenoviral vaccine or an mRNA vaccine or an inactivated vaccine or a DNA vaccine of the same virus, pathogen or tumor.

[0029] The present invention includes the full length sequences of said vaccine platform antigens as well as any truncated sequences, e.g., SEQ ID NO:76, SEQ ID NO:77, SEQ ID NO:78.

[0030] The present invention includes any possible variants of said fusion protein vaccine antigens, including mutant sequences such as naturally occurring point mutations / deletion mutations / truncations, any combination of naturally occurring point mutations, subtypes generated after mutation, and artificial point mutations / deletion mutations / truncations constructed by the inventors to enhance vaccine efficacy, e.g., SEQ ID NO: 79, SEQ ID NO: 80, SEQ ID NO: 81, SEQ ID NO: 82, SEQ ID NO: 83, SEQ ID NO: 84.

[0031] The invention also includes multivalent combination vaccines consisting of any of the vaccines of the invention as one component of a vaccine with another vaccine of the invention or with another vaccine different from the vaccine of the invention, e.g., another virus or pathogen or tumor vaccine, e.g., a multivalent vaccine in which the SARS-COV-2 fusion protein vaccine of the invention is combined with an influenza vaccine or other vaccine, e.g., any of the vaccines of the invention is combined with an adenoviral vaccine or an mRNA vaccine or an inactivated vaccine or a DNA vaccine of the same virus, pathogen or tumor in a sequential or simultaneous immunization procedure, e.g., a SARS-COV-2 fusion protein vaccine is combined with an adenoviral vaccine or an mRNA vaccine or an inactivated vaccine or a DNA vaccine of SARS-COV-2 ... may be immunized with the DNA vaccine in a sequential or simultaneous immunization procedure, for example, the order of immunization may be: 1) first with the SARS-COV-2 fusion protein vaccine of the present invention, then with the SARS-COV-2 adenovirus vaccine, mRNA vaccine, inactivated vaccine, or DNA vaccine; 2) first with the SARS-COV-2 adenovirus vaccine, mRNA vaccine, inactivated vaccine, or DNA vaccine, then with the SARS-COV-2 fusion protein vaccine; and 3) simultaneously with the SARS-COV-2 fusion protein vaccine and the SARS-COV-2 adenovirus vaccine, mRNA vaccine, inactivated vaccine, or DNA vaccine. When used in combination, the vaccines for combination use may be prepared as a convenient kit, as is known in the art.

[0032] Compared with the prior art, the present invention has the following beneficial effects, including but not limited to:

[0033] 1. In the IFN-tumor or viral antigen-immunoglobulin Fc (or antibody) vaccine platform provided by the present invention, the antigen can be varied in multiple components and can be a tumor-associated antigen or a virus-specific antigen, thereby improving the flexibility and scope of use of the vaccine platform.

[0034] 2. In the IFN-tumor or viral antigen-immunoglobulin Fc (or antibody) vaccine platform provided by the present invention, interferon (IFN) among them promotes the migration and maturation of antigen-presenting cells and increases the expression of costimulatory molecules, thereby favoring antigen presentation to T cells; at the same time, the Fc region (or antibody) of the vaccine platform, on the one hand, increases the molecular weight of the antigen and increases its half-life, and on the other hand, the Fc region (or antibody) binds to the Fc receptor on the surface of antigen-presenting cells, promoting the processing and presentation of the antigen by antigen-presenting cells, thereby favoring the production of an immune response.

[0035] 3. The IFN-tumor or viral antigen-immunoglobulin Fc (or antibody) vaccine platform provided by the present invention is expressed by eukaryotic HEK293 cell expression system, and the proteins expressed by HEK293 cells are close to natural protein molecules in terms of molecular structure, physical and chemical properties, protein modifications, and protein biological functions.

[0036] 4. The IFN-tumor or viral antigen-immunoglobulin Fc (or antibody) vaccine platform provided by the present invention has two structures, homodimer or heterodimer, and has better selection against different antigens.

[0037] 5. The IFN-tumor or viral antigen-immunoglobulin Fc vaccine platform provided by the present invention can activate DCs, enhance DC cross-presentation, and generate strong B cell and T cell immune responses by fusing a Th cell helper epitope, for example, a Pan epitope, utilizing a DC-targeting antibody such as anti-PD-L1, and adding various adjuvants that stimulate the immune response.

[0038] 6. The IFN-tumor or viral antigen-immunoglobulin Fc (or antibody) vaccine platform provided by the present invention has broad applications and can be used not only as a prophylactic vaccine but also as a therapeutic vaccine.

[0039] 7. The IFN-tumor or viral antigen-immunoglobulin Fc (or antibody) vaccine platform provided by the present invention can be used alone, but also in combination with existing HBsAg commercial vaccines, nucleoside / nucleotide analogues, as therapeutic vaccines.

[0040] 8. The vaccine provided by the present invention can be used as a component of a vaccine to form a multivalent combination vaccine with other viruses or pathogens or tumor vaccines.

[0041] 9. Any fusion protein vaccine in the vaccine platform provided by the present invention can be immunized with an adenovirus vaccine or an mRNA vaccine or an inactivated vaccine or a DNA vaccine of the same virus, pathogen, or tumor in a sequential or simultaneous immunization procedure.

[0042] 10. The present invention provides full length and any truncated sequences of vaccine platform antigens.

[0043] 11. The present invention provides any possible variants of vaccine platform antigens, including mutant sequences such as natural point mutations / deletion mutations / gain mutations / truncations, any combination of natural point mutations, subtypes generated after mutation, and artificial point mutations / deletion mutations / gain mutations / truncations constructed by the present inventors to enhance vaccine efficacy.

[0044] The sequence information related to the present invention is shown below. 1. Unit composition: SEQ ID NO: 1: Mouse mIFNα4 amino acid sequence (mIFNα) CDLPHTYNLGNKRALTVLEEMRRLPPLSCLKDRKDFGFPLEKVDNQQIQKAQAILVLRDLTQQILNLFTSKDLSATWNATLLDSFCNDLHQQLNDLKACVMQEPPLTQEDSLLAVRTYFHRITVYLRKKKHSLCAWEVIRAEVWRALSSSTNLLARLSEEKE SEQ ID NO: 21: Human IFNα2 amino acid sequence (hIFNα) CDLPQTHSLGSRRTLMLLAQMRRISLFSCLKDRHDFGFPQEEFGNQFQKAETIPVLHEMIQQIFNLFSTKDSSAAWDETLLDKFYTELYQQLNDLEACVIQGVGVTETPLMKEDSILAVRKYFQRITLYLKEKKYSPCAWEVVRAEIMRSFSLSTNLQESLRSKE SEQ ID NO.22: Human mutant IFNα2 (Q124R) amino acid sequence (hmIFNα) CDLPQTHSLGSRRTLMLLAQMRRISLFSCLKDRHDFGFPQEEFGNQFQKAETIPVLHEMIQQIFNLFSTKDSSAAWDETLLDKFYTELYQQLNDLEACVIQGVGVTETPLMKEDSILAVRKYFRRITLYLKEKKYSPCAWEVVRAEIMRSFSLSTNLQESLRSKE SEQ ID NO: 2: Human IgG1-Fc amino acid sequence EPKSCDKTHTCPPCPAPELLGGPSVFLFPPKPKDQLMISRTPEVTCVVVDVSHEDPEVKFNWYVDGVEVHNAKTKPREEQYNSTYRVVSVLTVLHQDWLNGKEYKCKVSNKALPAP IEKTISKAKGQPREPQVYTLPPSRDELTKNQVSLTCLVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFLYSKLTVDKSRWQQGNVFSCSVLHEALHNHYTQKSLSLSPGKHV SEQ ID NO: 23: Heterodimeric Fc-hole DKTHTCPPCPAPELLGGPSVFLFPPKPKDTLMISRTPEVTCVVVDVSHEDPEVKFNWYVDGVEVHNAKTKPREEQYNSTYRVVSVLTVLHQDWLNGKEYKCKVSNKALPAPIE KTISKAKGQPREPQVCTLPPSRDELTKNQVSLSCAVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLVSKLTVDKSRWQQGNVFSCSVMHEALHNHYTQKSLSLSPGK SEQ ID NO: 24: Heterodimeric Fc-knob DKTHTCPPCPAPELLGGPSVFLFPPKPKDTLMISRTPEVTCVVVDVSHEDPEVKFNWYVDGVEVHNAKTKPREEQYNSTYRVVSVLTVLHQDWLNGKEYKCKVSNKALPAPIE KTISKAKGQPREPQVYTLPPCRDELTKNQVSLWCLVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLYSKLTVDKSRWQQGNVFSCSVMHEALHNHYTQKSLSLSPGK SEQ ID NO: 3: Th helper epitope Pan HLA DR-binding epitope (PADER) amino acid sequence AKFVAAWTLKAAA SEQ ID NO: 4: Linker 1 amino acid sequence: GGGGSGGGGSGGGGS SEQ ID NO: 25: Linker 2 amino acid sequence: G.S.G.S.G. SEQ ID NO: 5: Signal peptide amino acid sequence: MARLCAFLMILVMMSYYWSACSLG SEQ ID NO: 6: Amino acid sequence of HBV Pres1 (ad subtype) MGGWSSKPRKGMGTNLSVPNPLGFFPDHQLDPAFGANSNNPDWDFNPIKDHWPAANQVGVGAFGPGLTPPHGGILGWSPQAQGILTTVSTIPPPASTNRQSGRQPTPSIPPLRDSHPQA SEQ ID NO: 26: Amino acid sequence of HBV Pres1 (ay subtype) MGQNLSTSNPLGFFPDHQLDPAFRANTANPDWDFNPNKDTWPDANKVGAGAFGLGFTPPHGGLLGWSPQAQGILQTLPANPPPASTNRQTGRQPTPLSPPLRNTHHPQA SEQ ID NO: 7: Amino acid sequence of HBV HBsAg (adr subtype) MENTTSGFLGPLLVLQAGFFLLTRILTIPQSLDSWWTSLNFLGGAPTCPGQNSQSPTSNHSPTSCPPICPGYRWMCLRRFIIFLFILLLCLIFLLVLLDYQGMLPVCPLLPGT STTSTGPCKTCTIPAQGTSMFPSCCCTKPSDGNCTCIPIPSSWAFARFLWEWASVRFSWLSLLVPFVQWFVGLSPTVWLSVIWMMWYWGPSLYNILSPFLPLLPIFFCLWVYI SEQ ID NO: 27: Amino acid sequence of HBV HBsAg (adw subtype) MENITSGLLGPLLVLQAGFFLLTRILTIPQSLDSWWTSLSFLGEAPVCLGQNSQSPTRNHSPTSCPPICPGYRWMCLRRFIIFLFILLLCLIFLLVLLDYQGMLPVCPLIPGS TTTSTGPCKTCTTPAQGNSMFPSCCCTKPTDGNCTCIPIPSSWAFAKYLWEWASVRFSWLSLLVPFVQWFVGLSPTVWLSAIWMIWYWGPSLYSIVCPFTPLLQIFCCLWVFI SEQ ID NO: 28: Amino acid sequence of HBV HBsAg (ayw subtype) MENITSGFLGPLLVLQAGFFLLTRILTIPQSLDSWWTSLNFLGGTTVCLGQSSQSPTSNHSPTSCPPTCPGYRWMCLRRFIIFLFILLLCLIFLLVLLDYQGMLPVCPLIPGS STTSTGPCRTCMTTAQGTSMYPSCCCTKPSDGNCTCIPIPSSWAFGKFLWEWASARFSWLSLLVPFVQWFVGLSPTVWLSVIWMMWYWGPSLYSILSPFLPLLPIFFCLWVYI SEQ ID NO: 8: Amino acid sequence of SARS-CoV-2 RBD RVQPTESIVRFPNITNLCPFGEVFNATRFASVYAWNRKRISNCVADYSVLYNSASFSTFKCYGVSPTKLNDLCFTNVYADSFVIRGDEVRQIAPGQTGKIADYNYKLPDDF TGCVIAWNSNNLDSKVGGNYNYLYRLFRKSNLKPFERDISTEIYQAGSTPCNGVEGFNCYFPLQSYGFQPTNGVGYQPYRVVVLSFELLHAPATVCGPKKSTNLVKNKCVNF SEQ ID NO: 9: Influenza virus HA amino acid sequence DTICIGYHANNSTDTVDTVLEKNVTVTHSVNLEDSHNGKLCRLKGIAPLQLGKCNIAGWLLGNPECDPLLPVRSWSYIVETPNSENGICYPGDFIDYEELREQLSSVSSFERFEIFPKESSWPNHNTNGVTAACSHEGKSSFYRNLLWLTEKEGSYPKLKNS YVNKKGKEVLVLWGIHPPNSKEQQNLYQNENAYVSVVTSNYNRRFTPEIAERPKVRDQAGRMNYYWTLLKPGDTIIFEANGNLIAPMYAFALSRGFGSGIITSNASMHECNTKCQTPLGAINSSLPYQNIHPVTIGECPKYVRSAKLRMVTGLRNNPSIQSR SEQ ID NO: 10: Amino acid sequence of HPV-E7 antigen MHGDTPTLHEYMLDLQPETTDLYCYEQLNDSSEEEDEIDGPAGQAEPDRAHYNIVTFCCKCDSTLRLCVQSTHVDIRTLEDLLMGTLGIVCPICSQKP 2. Murine-derived IFN vaccine mIFNα-antigen-Fc sequence: SEQ ID NO: 11: mIFNα-Pres1-Fc amino acid sequence in homodimer CDLPHTYNLGNKRALTVLEEMRRLPPLSCLKDRKDFGFPLEKVDNQQIQKAQAILVLRDLTQQILNLFTSKDLSATWNATLLDSFCNDLHQQLNDLKACVMQEPPLTQEDSLLAVRTYFHRITVYLRKKKHSLCAWEVIRAEVWRALSSSTNLLARLSEEKESGGGGSGGGGSGGGGSGGGGRTMGQNLSTSNPLGFFPDHQLDPAFRANTANPDWDFNPNKDTWPDANKVGAGAFGLGFTPPHGGLLGWSPQAQGILQTLPANPPPASTNRQTGRQPTPLSPPLRNTHPQAFEEPKSCDKTHTCPPCPAPELLGGPSVFLFPPKPKDQLMISRTPEVTCVVVDVSHEDPEVKFNWYVDGVEVHNAKTKPREEQYNSTYRVVSVLTVLHQDWLNGKEYKCKVSNKALPAPIEKTISKAKGQPREPQVYTLPPSRDELTKNQVSLTCLVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFLYSKLTVDKSRWQQGNVFSCSVLHEALHNHYTQKSLSLSPGKHV SEQ ID NO: 12: Amino acid sequence of mIFNα-RBD(SARS-CoV-2)-Fc in homodimer CDLPHTYNLGNKRALTVLEEMRRLPPLSCLKDRKDFGFPLEKVDNQQIQKAQAILVLRDLTQQILNLFTSKDLSATWNATLLDSFCNDLHQQLNDLKACVMQEPPLTQEDSLLAVRTYFHRITVYLRKKKHSLCAWEVIRAEVWRALSSSTNLLARLSEE KESGGGGSGGGGSGGGGSGGGGRTRVQPTESIVRFPNITNLCPFGEVFNATRFASVYAWNRKRISNCVADYSVLYNSASFSTFKCYGVSPTKLNDLCFTNVYADSFVIRGDEVRQIAPGQTGKIADYNYKLPDDFTGCVIAWNSNNLDSKVGGNYNYLYRL FRKSNLKPFERDISTEIYQAGSTPCNGVEGFNCYFPLQSYGFQPTNGVGYQPYRVVVLSFELLHAPATVCGPKKSTNLVKNKCVNFFEEPKSCDKTHTCPPCPAPELLGGPSVFLFPPKPKDQLMISRTPEVTCVVDVSHEDPEVKFNWYVDGVEVHNA KTKPREEQYNSTYRVVSVLTVLHQDWLNGKEYKCKVSNKALPAPIEKTISKAKGQPREPQVYTLPPSRDELTKNQVSLTCLVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFLYSKLTVDKSRWQQGNVFSCSVLHEALHNHYTQKSLSLSPGKHV SEQ ID NO: 13: mIFNα-HA-Fc amino acid sequence in homodimer CDLPHTYNLGNKRALTVLEEMRRLPPLSCLKDRKDFGFPLEKVDNQQIQKAQAILVLRDLTQQILNLFTSKDLSATWNATLLDSFCNDLHQQLNDLKACVMQEPPLTQEDSLLAVRTYFHRITVYLRKKKHSLCAWEVIRAEVWRALSSSTNLLARLSEEKESGGGGSGGGGSGGGGSGGGGRTDT ICIGYHANNSTDTVDTVLEKNVTVTHSVNLLEDSHNGKLCRLKGIAPLQLGKCNIAGWLLGNPECDPLLPVRSWSYIVETPNSENGICYPGDFIDYEELREQLSSVSSFERFEIFPKESSWPNHNTNGVTAACSHEGKSSFYRNLLWLTEKEGSYPKLKNSYVNKKGKEVLVLWGIHHPPNSKEQQ NLYQNENAYVSVVTSNYNRRFTPEIAERPKVRDQAGRMNYYWTLLKPGDTIIFEANGNLIAPMYAFALSRGFGSGIITSNASMHECNTKCQTPLGAINSSLPYQNIHPVTIGECPKYVRSAKLRMVTGLRNNPSIQSRFEEPKSCDKTHTCPPCPAPELLGGPSVFLFPPKDQLMISRTPEVTC VVVDVSHEDPEVKFNWYVDGVEVHNAKTKPREEQYNSTYRVVSVLTVLHQDWLNGKEYKCKVSNKALAPIEKTISKAKGQPREPQVYTLPPS RDELTKNQVSLTCLVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFLYSKLTVDKSRWQQGNVFSCSVLHEALHNHYTQKSLSLSPGKHV SEQ ID NO: 14: mIFNα-E7(HPV)-Fc amino acid sequence in homodimer CDLPHTYNLGNKRALTVLEEMRRLPPLSCLKDRKDFGFPLEKVDNQQIQKAQAILVLRDLTQQILNLFTSKDLSATWNATLLDSFCNDLHQQLNDLKACVMQEPPLTQEDSLLAVRTYFHRITVYLRKKKHSLCAWEVIRAEVWRALSSSTNLLARLSEEKESGGGGSGGGGSGGGGSGGGGRTMHGDTPTLHEYMLDLQPETTDLYCYEQLNDSSEEEDEIDGPAGQAEPDRAHYNIVTFCCKCDSTLRLCVQSTHVDIRTLEDLLMGTLGIVCPICSQKPFEEPKSCDKTHTCPPCPAPELLGGPSVFLFPPKPKDQLMISRTPEVTCVVVDVSHEDPEVKFNWYVDGVEVHNAKTKPREEQYNSTYRVVSVLTVLHQDWLNGKEYKCKVSNKALPAPIEKTISKAKGQPREPQVYTLPPSRDELTKNQVSLTCLVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFLYSKLTVDKSRWQQGNVFSCSVLHEALHNHYTQKSLSLSPGKHV SEQ ID NO: 15: Amino acid sequence of the first chain mIFNα-Fc-hole in a heterodimer CDLPHTYNLGNKRALTVLEEMRRLPPLSCLKDRKDFGFPLEKVDNQQIQKAQAILVLRDLTQQILNLFTSKDLSATWNATLLDSFCNDLHQQLNDLKACVMQEPPLTQEDSLLAVRTYFHRITVYLRKKKHSLCAWEVIRAEVWRALSSSTNLLARLSEEKESGGGGSGGGGSGGGGSGGGGRTDKTHTCPPCPAPELLGGPSVFLFPPKPKDTLMISRTPEVTCVVVDVSHEDPEVKFNWYVDGVEVHNAKTKPREEQYNSTYRVVSVLTVLHQDWLNGKEYKCKVSNKALPAPIEKTISKAKGQPREPQVCTLPPSRDELTKNQVSLSCAVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLVSKLTVDKSRWQQGNVFSCSVMHEALHNHYTQKSLSLSPGK SEQ ID NO: 16: Amino acid sequence of the second chain Pres1-Fc-knob in the heterodimeric mIFNα-Pres1-Fc MGQNLSTSNPLGFFPDHQLDPAFRANTANPDWDFNPNKDTWPDANKVGAGAFGLGFTPPHGGLLGWSPQAQGILQTLPANPPPASTNRQTGRQPTPLSPPLRNTHPQAFEDKTHTCPPCPAPELLGGPSVFLFPPKPKDTLMISRTPEVTCVVVDVSHEDPEVKFNWY VDGVEVHNAKTKPREEQYNSTYRVVSVLTVLHQDWLNGKEYKCKVSNKALPAPIEKTISKAKGQPREPQVYTLPPCRDELTKNQVSLWCLVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLYSKLTVDKSRWQQGNVFSCSVMHEALHNHYTQKSLSLSPGK SEQ ID NO: 17: Amino acid sequence of the second chain RBD(SARS-CoV-2)-Fc-knob in the heterodimeric mIFNα-RBD(SARS-CoV-2)-Fc RVQPTESIVRFPNITNLCPFGEVFNATRFASVYAWNRKRISNCVADYSVLYNSASFSTFKCYGVSPTKLNDLCFTNVYADSFVIRGDEVRQIAPGQTGKIADYNYKLPDDFTG CVIAWNSNNLDSKVGGNYNYLYRLFRKSNLKPFERDISTEIYQAGSTPCNGVEGFNCYFPLQSYGFQPTNGVGYQPYRVVVLSFELLHAPATVCGPKKSTNLVKNKCVNFFED KTHTCPPCPAPELLGGPSVFLFPPKPKDTLMISRTPEVTCVVVDVSHEDPEVKFNWYVDGVEVHNAKTKPREEQYNSTYRVVSVLTVLHQDWLNGKEYKCKVSNKALPAPIEK TISKAKGQPREPQVYTLPPCRDELTKNQVSLWCLVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLYSKLTVDKSRWQQGNVFSCSVMHEALHNHYTQKSLSLSPGK SEQ ID NO: 18: Amino acid sequence of the second chain HA-Fc-knob in the heterodimeric mIFNα-HA-Fc DTICIGYHANNSTDTVDTVLEKNVTVTHSVNLLEDSHNGKLCRLKGIAPLQLGKCNIAGWLLGNPECDPLLPVRSWSYIVETPNSENGICYPGDFIDYEELREQLSSVSSFERFEIFPKESSWPNHNTNGVTAACSHE GKSSFYRNLLWLTEKEGSYPKLKNSYVNKKGKEVLVLWGIHPPNSKEQQNLYQNENAYVSVVTSNYNRRFTPEIAERPKVRDQAGRMNYYWTLLKPGDTIIFEANGNLIAPMYAFALSRGGFGSGIITSNASMHECNTK CQTPLGAINSSLPYQNIHPVTIGECPKYVRSAKLRMVTGLRNNPSIQSRFEDKTHTCPPCPAPELLGGPSVFLFPPKPKDTLMISRTPEVTCVVDVSHEDPEVKFNWYVDGVEVHNAKTKPREEQYNSTYRVVSVLTV LHQDWLNGKEYKCKVSNKALPAPIEKTISKAKGQPREPQVYTLPPCRDELTKNQVSLWCLVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLYSKLTVDKSRWQQGNVFSCSVMHEALHNHYTQKSLSLSPGK SEQ ID NO: 19: Amino acid sequence of the second chain E7-Fc-knob in the heterodimeric mIFNα-E7(HPV)-Fc MHGDTPTLHEYMLDLQPETTDLYCYEQLNDSSEEEDEIDGPAGQAEPDRAHYNIVTFCCKCDSTLRLCVQSTHVDIRTLEDLLMGTLGIVCPICSQKPFEDKTHTCPPCPAPELLGGPSVFLFPPKPKDTLMISRTPEVTCVVDVSHEDPEVKFNWYVDGVE VHNAKTKPREEQYNSTYRVVSVLTVLHQDWLNGKEYKCKVSNKALPAPIEKTISKAKGQPREPQVYTLPPCRDELTKNQVSLWCLVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLYSKLTVDKSRWQQGNVFSCSVMHEALHNHYTQKSLSLSPGK 3. Vaccine IFNα-Pan-Antigen-Fc sequence containing the Pan epitope of murine IFN: SEQ ID NO: 29: mIFNα-Pan-Pres1-Fc amino acid sequence in homodimer CDLPHTYNLGNKRALTVLEEMRRLPPLSCLKDRKDFGFPLEKVDNQQIQKAQAILVLRDLTQQILNLFTSKDLSATWNATLLDSFCNDLHQQLNDLKACVMQEPPLTQEDSLLAVRTYFHRITVYLRKKKHSL CAWEVIRAEVWRALSSSTNLLARLSEEKEGGGGSGGGGSGGGGSRTAKFVAAWTLKAAAGSGSGSMGQNLSTSNPLGFFPDHQLDPAFRANTANPDWDFNPNKDTWPDANKVGAGAFGLGFTPPHGGLLGWSPQ AQGILQTLPANPPPASTNRQTGRQPTPLSPPLRNTHPQAFEDKTHTCPPCPAPELLGGPSVFLFPPKPKDQLMISRTPEVTCVVVDVSHEDPEVKFNWYVDGVEVHNAKTKPREEQYNSTYRVVSVLTVLHQD WLNGKEYKCKVSNKALAPIEKTISKAKGQPREPQVYTLPPSRDELTKNQVSLTCLVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFLYSKLTVDKSRWQQGNVFSCSVLHEALHNHYTQKSLSLSPGK SEQ ID NO: 30: mIFNα-Pan-RBD(SARS-CoV-2)-Fc amino acid sequence in homodimer CDLPHTYNLGNKRALTVLEEMRRLPPLSCLKDRKDFGFPLEKVDNQQIQKAQAILVLRDLTQQILNLFTSKDLSATWNATLLDSFCNDLHQQLNDLKACVMQEPPLTQEDSLLAVRTYFHRITVYLRKKKHSLCAWEVIRAEVWRALSSSTNLLARLSEEKE GGGGSGGGGSGGGGSRTAKFVAAWTLKAAAGSGSGSRVQPTESIVRFPNITNLCPFGEVFNATRFASVYAWNRKRISNCVADYSVLYNSASFSTFKCYGVSPTKLNDLCFTNVYADSFVIRGDEVRQIAPGQTGKIADYNYKLPDDFTGCVIAWNSNNLDSK VGGNYNYLYRLFRKSNLKPFERDISTEIYQAGSTPCNGVEGFNCYFPLQSYGFQPTNGVGYQPYRVVVLSFELLHAPATVCGPKKSTNLVKNKCVNFFEDKTHTCPPCPAPELLGGPSVFLFPPKPKDQLMISRTPEVTCVVVDVSHEDPEVKFNWYVDGVE VHNAKTKPREEQYNSTYRVVSVLTVLHQDWLNGKEYKCKVSNKALPAPIEKTISKAKGQPREPQVYTLPPSRDELTKNQVSLTCLVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFLYSKLTVDKSRWQQGNVFSCSVLHEALHNHYTQKSLSLSPGK SEQ ID NO: 31: mIFNα-Pan-HA-Fc amino acid sequence in homodimer CDLPHTYNLGNKRALTVLEEMRRLPPLSCLKDRKDFGFPLEKVDNQQIQKAQAILVLRDLTQQILNLFTSKDLSATWNATLLDSFCNDLHQQLNDLKACVMQEPPLTQEDSLLAVRTYFHRITVYLRKKKHSLCAWEVIRAEVWRALSSSTNLLARLSEEKEGGGGGSGGGGSGGGGSRTAKFVAAWTL KAAAGSGSGSDTICIGYHANNSTDTVDTVLEKNVTVTHSVNLLEDSHNGKLCRLKGIAPLQLGKCNIAGWLLGNPECDPLLPVRSWSYIVETPNSENGICYPGDFIDYEELREQLSSVSSFERFEIFPKESSWPNHNTNGVTAACSHEGKSSFYRNLLWLTEKEGSYPKLKNSYVNKKGKEVLVLWGI HHPPNSKEQQNLYQNENAYVSVVTSNYNRRFTPEIAERPKVRDQAGRMNYYWTLLKPGDTIIFEANGNLIAPMYAFALSRGFGSGIITSNASMHECNTKCQTPLGAINSSLPYQNIHPVTIGECPKYVRSAKLRMVTGLRNNPSIQSRFEDKTHTCPPCPAPELLGGPSVFLFPPKPKDQLMISRTPE VTCVVVDVSHEDPEVKFNWYVDGVEVHNAKTKPREEQYNSTYRVVSVLTVLHQDWLNGKEYKCKVSNKALPAPIEKTISKAKGQPREPQVYTLP PSRDELTKNQVSLTCLVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFLYSKLTVDKSRWQQGNVFSCSVLHEALHNHYTQKSLSLSPGK SEQ ID NO: 32: mIFNα-Pan-E7(HPV)-Fc amino acid sequence in homodimer CDLPHTYNLGNKRALTVLEEMRRLPPLSCLKDRKDFGFPLEKVDNQQIQKAQAILVLRDLTQQILNLFTSKDLSATWNATLLDSFCNDLHQQLNDLKACVMQEPPLTQEDSLLAVRTYFHRITVYLRKKKH SLCAWEVIRAEVWRALSSSTNLLARLSEEKEGGGGSGGGGSGGGGSRTAKFVAAWTLKAAAGSGSGSMHGDTPTLHEYMLDLQPETTDLYCYEQLNDSSEEEDEIDGPAGQAEPDRAHYNIVTFCCKCDST LRLCVQSTHVDIRTLEDLLMGTLGIVCPICSQKPFEDKTHTCPPCPAPELLGGPSVFLFPPKPKDQLMISRTPEVTCVVVDVSHEDPEVKFNWYVDGVEVHNAKTKPREEQYNSTYRVVSVLTVLHQDWLN GKEYKCKVSNKALAPIEKTISKAKGQPREPQVYTLPPSRDELTKNQVSLTCLVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFLYSKLTVDKSRWQQGNVFSCSVLHEALHNHYTQKSLSLSPGK SEQ ID NO: 33: First chain mIFNα-Fc-hole amino acid sequence in the heterodimer CDLPHTYNLGNKRALTVLEEMRRLPPLSCLKDRKDFGFPLEKVDNQQIQKAQAILVLRDLTQQILNLFTSKDLSATWNATLLDSFCNDLHQQLNDLKACVMQEPPLTQEDSLLAVRTYFHRITVYLRKKKHSLCAWEVIRAEVWRALSSSTNLLARLSEEKESGGGGSGGGGSGGGGSGGGGRTDKTHTCPPCPAPELLGGPSVF LFPPKPKDTLMISRTPEVTCVVVDVSHEDPEVKFNWYVDGVEVHNAKTKPREEQYNSTYRVVSVLTVLHQDWLNGKEYKCKVSNKALPAPIEKTISKAKGQPR EPQVCTLPPSRDELTKNQVSLSCAVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLVSKLTVDKSRWQQGNVFSCSVMHEALHNHYTQKSLSLSPGK SEQ ID NO: 34: Amino acid sequence of the second chain Pan-Pres1-Fc-knob in the heterodimeric mIFN-Pan-Pres1-Fc AKFVAAWTLKAAAGSGSGSMGQNLSTSNPLGFFPDHQLDPAFRANTANPDWDFNPNKDTWPDANKVGAGAFGLGFTPPHGGLLGWSPQAQGILQTLPANPPPASTNRQTGRQPTPLSPPLRNTHPQAFEDKTHTCPPCPAPELLGGPSVFLFPPKPKDTLMISRTPEVTCVVVDVSHE DPEVKFNWYVDGVEVHNAKTKPREEQYNSTYRVVSVLTVLHQDWLNGKEYKCKVSNKALPAPIEKTISKAKGQPREPQVYTLPPCRDELTKNQVSLWCLVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLYSKLTVDKSRWQQGNVFSCSVMHEALHNHYTQKSLSLSPGK SEQ ID NO: 35: Amino acid sequence of the second chain Pan-RBD(SARS-CoV-2)-Fc-knob in the heterodimeric mIFNα-Pan-RBD(SARS-CoV-2)-Fc AKFVAAWTLKAAAGSGSGSRVQPTESIVRFPNITNLCPFGEVFNATRFASVYAWNRKRISNCVADYSVLYNSASFSTFKCYGVSPTKLNDLCFTNVYADSFVIRGDEVRQIAPGQTG KIADYNYKLPDDFTGCVIAWNSNNLDSKVGGNYNYLYRLFRKSNLKPFERDISTEIYQAGSTPCNGVEGFNCYFPLQSYGFQPTNGVGYQPYRVVVLSFELLHAPATVCGPKKSTNLV KNKCVNFFEDKTHTCPPCPAPELLGGPSVFLFPPKPKDTLMISRTPEVTCVVVDVSHEDPEVKFNWYVDGVEVHNAKTKPREEQYNSTYRVVSVLTVLHQDWLNGKEYKCKVSNKALP APIEKTISKAKGQPREPQVYTLPPCRDELTKNQVSLWCLVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLYSKLTVDKSRWQQGNVFSCSVMHEALHNHYTQKSLSLSPGK SEQ ID NO: 36: Amino acid sequence of the second chain Pan-HA-Fc-knob in the heterodimeric mIFNα-Pan-HA-Fc AKFVAAWTLKAAAGSGSGSDTICIGYHANNSTDTVDTVLEKNVTVTHSVNLLEDSHNGKLCRLKGIAPLQLGKCNIAGWLLGNPECDPLLPVRSWSYIVETPNSENGICYPGDFIDYEELREQLSSVSSFERFEIFPKESSWP NHNTNGVTAACSHEGKSSFYRNLLWLTEKEGSYPKLKNSYVNKKGKEVLVLWGIHHPPNSKEQQNLYQNENAYVSVVTSNYNRRFTPEIAERPKVRDQAGRMNYYWTLLKPGDTIIFEANGNLIAPMYAFALSRGGFGSGIITSN ASMHECNTKCQTPLGAINSSLPYQNIHPVTIGECPKYVRSAKLRMVTGLRNNPSIQSRFEDKTHTCPPCPAPELLGGPSVFLFPPKPKDTLMISRTPEVTCVVVDVSHEDPEVKFNWYVDGVEVHNAKTKPREEQYNSTYRVV SVLTVLHQDWLNGKEYKCKVSNKALPAPIEKTISKAKGQPREPQVYTLPPCRDELTKNQVSLWCLVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLYSKLTVDKSRWQQGNVFSCSVMHEALHNHYTQKSLSLSPGK SEQ ID NO: 37: Amino acid sequence of the second chain Pan-E7-Fc-knob in the heterodimeric mIFNα-Pan-E7(HPV)-Fc AKFVAAWTLKAAAGSGSGSMHGDTPTLHEYMLDLQPETTDLYCYEQLNDSSEEEDEIDGPAGQAEPDRAHYNIVTFCCKCDSTLRLCVQSTHVDIRTLEDLLMGTLGIVCPICSQKPFEDKTHTCPPCPAPELLGGPSVFLFPPKPKDTLMISRTPEVTCVVVDVSHEDPEVK FNWYVDGVEVHNAKTKPREEQYNSTYRVVSVLTVLHQDWLNGKEYKCKVSNKALPAPIEKTISKAKGQPREPQVYTLPPCRDELTKNQVSLWCLVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLYSKLTVDKSRWQQGNVFSCSVMHEALHNHYTQKSLSLSPGK 4. Human IFN Vaccine hIFNα-Antigen-Fc Sequence: SEQ ID NO: 38: hIFNα-Pres1-Fc amino acid sequence in homodimer CDLPQTHSLGSRRTLMLLAQMRRISLFSCLKDRHDFGFPQEEFGNQFQKAETIPVLHEMIQQIFNLFSTKDSSAAWDETLLDKFYTELYQQLNDLEACVIQGVGVTETPLMKEDSILAVRKYFQRITLY LKEKKYSPCAWEVVRAEIMRSFSLSTNLQESLRSKEGGGGSGGGGSGGGGSRTMGQNLSTSNPLGFFPDHQLDPAFRANTANPDWDFNPNKDTWPDANKVGAGAFGLGFTPPHGGLLGWSPQAQGILQTL PANPPPASTNRQTGRQPTPLSPPLRNTHPQAFEDKTHTCPPCPAPELLGGPSVFLFPPKPKDQLMISRTPEVTCVVVDVSHEDPEVKFNWYVDGVEVHNAKTKPREEQYNSTYRVVSVLTVLHQDWLNG KEYKCKVSNKALAPIEKTISKAKGQPREPQVYTLPPSRDELTKNQVSLTCLVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFLYSKLTVDKSRWQQGNVFSCSVLHEALHNHYTQKSLSLSPGK SEQ ID NO: 39: hIFNα-RBD (SARS-CoV-2)-Fc amino acid sequence in homodimer CDLPQTHSLGSRRTLMLLAQMRRISLFSCLKDRHDFGFPQEEFGNQFQKAETIPVLHEMIQQIFNLFSTKDSSAAWDETLLDKFYTELYQQLNDLEACVIQGVGVTETPLMKEDSILAVRKYFQRITLYLKEKKYSPCAWEVVRAEIMRSFSLSTNLQ ESLRSKEGGGGSGGGGSGGGGSRTRVQPTESIVRFPNITNLCPFGEVFNATRFASVYAWNRKRISNCVADYSVLYNSASFSTFKCYGVSPTKLNDLCFTNVYADSFVIRGDEVRQIAPGQTGKIADYNYKLPDDFTGCVIAWNSNNLDSKVGGNYNYL YRLFRKSNLKPFERDISTEIYQAGSTPCNGVEGFNCYFPLQSYGFQPTNGVGYQPYRVVVLSFELLHAPATVCGPKKSTNLVKNKCVNFFEDKTHTCPPCPAPELLGGPSVFLFPPKPKDQLMISRTPEVTCVVVDVSHEDPEVKFNWYVDGVEVHNA KTKPREEQYNSTYRVVSVLTVLHQDWLNGKEYKCKVSNKALPAPIEKTISKAKGQPREPQVYTLPPSRDELTKNQVSLTCLVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFLYSKLTVDKSRWQQGNVFSCSVLHEALHNHYTQKSLSLSPGK SEQ ID NO: 40: hIFNα-HA-Fc amino acid sequence in homodimer CDLPQTHSLGSRRTLMLLAQMRRISLFSCLKDRHDFGFPQEEFGNQFQKAETIPVLHEMIQQIFNLFSTKDSSAAWDETLLDKFYTELYQQLNDLEACVIQGVGVTETPLMKEDSILAVRKYFQRITLYLKEKKYSPCAWEVVRAEIMRSFSLSTNLQESLRSKEGGGGSGGGGSGGGGSRTDT ICIGYHANNSTDTVDTVLEKNVTVTHSVNLLEDSHNGKLCRLKGIAPLQLGKCNIAGWLLGNPECDPLLPVRSWSYIVETPNSENGICYPGDFIDYEELREQLSSVSSFERFEIFPKESSWPNHNTNGVTAACSHEGKSSFYRNLLWLTEKEGSYPKLKNSYVNKKGKEVLVLWGIHHPPNSKE QQNLYQNENAYVSVVTSNYNRRFTPEIAERPKVRDQAGRMNYYWTLLKPGDTIIFEANGNLIAPMYAFALSRGFGSGIITSNASMHECNTKCQTPLGAINSSLPYQNIHPVTIGECPKYVRSAKLRMVTGLRNNPSIQSRFEDKTHTCPPCPAPELLGGPSVFLFPPKKDQLMISRTPEVTCV VVDVSHEDPEVKFNWYVDGVEVHNAKTKPREEQYNSTYRVVSVLTVLHQDWLNGKEYKCKVSNKALPAPIEKTISKAKGQPREPQVYTLPPS RDELTKNQVSLTCLVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFLYSKLTVDKSRWQQGNVFSCSVLHEALHNHYTQKSLSLSPGK SEQ ID NO: 41: hIFNα-E7(HPV)-Fc amino acid sequence in homodimer CDLPQTHSLGSRRTLMLLAQMRRISLFSCLKDRHDFGFPQEEFGNQFQKAETIPVLHEMIQQIFNLFSTKDSSAAWDETLLDKFYTELYQQLNDLEACVIQGVGVTETPLMKEDSILAVRKYFQRITLYLKEKKYSPCAWEVVRAEIMRSFSLSTNLQESLRSKEGGGGSGGGGSGGGGSRTMHGDTPTLHEYMLDLQPETTDLYCYEQLNDSSEEEDEIDGPAGQAEPDRAHYNIVTFCCKCDSTLRLCVQSTHVDIRTLEDLLMGTLGIVCPICSQKPFEDKTHTCPPCPAPELLGGPSVFLFPPKPKDQLMISRTPEVTCVVVDVSHEDPEVKFNWYVDGVEVHNAKTKPREEQYNSTYRVVSVLTVLHQDWLNGKEYKCKVSNKALPAPIEKTISKAKGQPREPQVYTLPPSRDELTKNQVSLTCLVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFLYSKLTVDKSRWQQGNVFSCSVLHEALHNHYTQKSLSLSPGK SEQ ID NO: 42: Amino acid sequence of the first chain hIFN-Fc-hole in the heterodimer CDLPQTHSLGSRRTLMLLAQMRRISLFSCLKDRHDFGFPQEEFGNQFQKAETIPVLHEMIQQIFNLFSTKDSSAAWDETLLDKFYTELYQQLNDLEACVIQGVGVTETPLMKEDSILAVRKYFQRITLYLKEKKYSPCAWEVVRAEIMRSFSLSTNLQESLRSKESGGGGSGGGGSGGGGSGGGGRTDKTHTCPPCPAPELLGGPSVFLFPPKPKDTLMISRTPEVTCVVVDVSHEDPEVKFNWYVDGVEVHNAKTKPREEQYNSTYRVVSVLTVLHQDWLNGKEYKCKVSNKALPAPIEKTISKAKGQPREPQVCTLPPSRDELTKNQVSLSCAVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFKLVSKLTVDKSRWQQGNVFSCSVMHEALHNHYTQKSLSLSPGK SEQ ID NO: 43: Amino acid sequence of the second chain Pres1-Fc-knob in the heterodimeric hIFNα-Pres1-Fc MGQNLSTSNPLGFFPDHQLDPAFRANTANPDWDFNPNKDTWPDANKVGAGAFGLGFTPPHGGLLGWSPQAQGILQTLPANPPPASTNRQTGRQPTPLSPPLRNTHPQAFEDKTHTCPPCPAPELLGGPSVFLFPPKPKDTLMISRTPEVTCVVVDVSHEDPEVKFNWY VDGVEVHNAKTKPREEQYNSTYRVVSVLTVLHQDWLNGKEYKCKVSNKALPAPIEKTISKAKGQPREPQVCTLPPSRDELTKNQVSLSCAVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLVSKLTVDKSRWQQGNVFSCSVMHEALHNHYTQKSLSLSPGK SEQ ID NO: 44: Second chain RBD(SARS-CoV-2)-Fc-knob amino acid sequence in heterodimeric hIFNα-RBD(SARS-CoV-2)-Fc RVQPTESIVRFPNITNLCPFGEVFNATRFASVYAWNRKRISNCVADYSVLYNSASFSTFKCYGVSPTKLNDLCFTNVYADSFVIRGDEVRQIAPGQTGKIADYNYKLPDDFTG CVIAWNSNNLDSKVGGNYNYLYRLFRKSNLKPFERDISTEIYQAGSTPCNGVEGFNCYFPLQSYGFQPTNGVGYQPYRVVVLSFELLHAPATVCGPKKSTNLVKNKCVNFFED KTHTCPPCPAPELLGGPSVFLFPPKPKDTLMISRTPEVTCVVVDVSHEDPEVKFNWYVDGVEVHNAKTKPREEQYNSTYRVVSVLTVLHQDWLNGKEYKCKVSNKALPAPIEK TISKAKGQPREPQVYTLPPCRDELTKNQVSLWCLVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLYSKLTVDKSRWQQGNVFSCSVMHEALHNHYTQKSLSLSPGK SEQ ID NO: 45: Amino acid sequence of the second chain HA-Fc-knob in heterodimeric hIFNα-HA-Fc DTICIGYHANNSTDTVDTVLEKNVTVTHSVNLLEDSHNGKLCRLKGIAPLQLGKCNIAGWLLGNPECDPLLPVRSWSYIVETPNSENGICYPGDFIDYEELREQLSSVSSFERFEIFPKESSWPNHNTNGVTAACSHE GKSSFYRNLLWLTEKEGSYPKLKNSYVNKKGKEVLVLWGIHPPNSKEQQNLYQNENAYVSVVTSNYNRRFTPEIAERPKVRDQAGRMNYYWTLLKPGDTIIFEANGNLIAPMYAFALSRGGFGSGIITSNASMHECNTK CQTPLGAINSSLPYQNIHPVTIGECPKYVRSAKLRMVTGLRNNPSIQSRFEDKTHTCPPCPAPELLGGPSVFLFPPKPKDTLMISRTPEVTCVVDVSHEDPEVKFNWYVDGVEVHNAKTKPREEQYNSTYRVVSVLTV LHQDWLNGKEYKCKVSNKALPAPIEKTISKAKGQPREPQVYTLPPCRDELTKNQVSLWCLVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLYSKLTVDKSRWQQGNVFSCSVMHEALHNHYTQKSLSLSPGK SEQ ID NO: 46: Amino acid sequence of the second chain E7(HPV)-Fc-knob in the heterodimeric hIFNα-E7(HPV)-Fc MHGDTPTLHEYMLDLQPETTDLYCYEQLNDSSEEEDEIDGPAGQAEPDRAHYNIVTFCCKCDSTLRLCVQSTHVDIRTLEDLLMGTLGIVCPICSQKPFEDKTHTCPPCPAPELLGGPSVFLFPPKPKDTLMISRTPEVTCVVDVSHEDPEVKFNWYVDGVE VHNAKTKPREEQYNSTYRVVSVLTVLHQDWLNGKEYKCKVSNKALPAPIEKTISKAKGQPREPQVYTLPPCRDELTKNQVSLWCLVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLYSKLTVDKSRWQQGNVFSCSVMHEALHNHYTQKSLSLSPGK 5. Vaccine containing the Pan epitope of human IFN α-Pan-Antigen-Fc sequence: SEQ ID NO: 47: hIFNα-Pan-Pres1-Fc amino acid sequence in homodimer CDLPQTHSLGSRRTLMLLAQMRRISLFSCLKDRHDFGFPQEEFGNQFQKAETIPVLHEMIQQIFNLFSTKDSSAAWDETLLDKFYTELYQQLNDLEACVIQGVGVTETPLMKEDSILAVRKYFQRITLYL KEKKYSPCAWEVVRAEIMRSFSLSTNLQESLRSKEAKFVAAWTLKAAAGSGSGSMGQNLSTSNPLGFFPDHQLDPAFRANTANPDWDFNPNKDTWPDANKVGAGAFGLGFTPPHGGLLGWSPQAQGILQT LPANPPPASTNRQTGRQPTPLSPPLRNTHPQAFEDKTHTCPPCPAPELLGGPSVFLFPPKPKDQLMISRTPEVTCVVVDVSHEDPEVKFNWYVDGVEVHNAKTKPREEQYNSTYRVVSVLTVLHQDWLNG KEYKCKVSNKALAPIEKTISKAKGQPREPQVYTLPPSRDELTKNQVSLTCLVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFLYSKLTVDKSRWQQGNVFSCSVLHEALHNHYTQKSLSLSPGK SEQ ID NO: 48: hIFNα-Pan-RBD(SARS-CoV-2)-Fc amino acid sequence in homodimer CDLPQTHSLGSRRTLMLLAQMRRISLFSCLKDRHDFGFPQEEFGNQFQKAETIPVLHEMIQQIFNLFSTKDSSAAWDETLLDKFYTELYQQLNDLEACVIQGVGVTETPLMKEDSILAVRKYFQRITLYLKEKKYSPCAWEVVRAEIMRSFSLSTNLQ ESLRSKEAKFVAAWTLKAAAGSGSGSRVQPTESIVRFPNITNLCPFGEVFNATRFASVYAWNRKRISNCVADYSVLYNSASFSTFKCYGVSPTKLNDLCFTNVYADSFVIRGDEVRQIAPGQTGKIADYNYKLPDDFTGCVIAWNSNNLDSKVGGNYNY LYRLFRKSNLKPFERDISTEIYQAGSTPCNGVEGFNCYFPLQSYGFQPTNGVGYQPYRVVVLSFELLHAPATVCGPKKSTNLVKNKCVNFFEDKTHTCPPCPAPELLGGPSVFLFPPKPKDQLMISRTPEVTCVVDVSHEDPEVKFNWYVDGVEVHNA KTKPREEQYNSTYRVVSVLTVLHQDWLNGKEYKCKVSNKALPAPIEKTISKAKGQPREPQVYTLPPSRDELTKNQVSLTCLVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFLYSKLTVDKSRWQQGNVFSCSVLHEALHNHYTQKSLSLSPGK SEQ ID NO: 49: hIFNα-Pan-HA-Fc amino acid sequence in homodimer CDLPQTHSLGSRRTLMLLAQMRRISLFSCLKDRHDFGFPQEEFGNQFQKAETIPVLHEMIQQIFNLFSTKDSSAAWDETLLDKFYTELYQQLNDLEACVIQGVGVTETPLMKEDSILAVRKYFQRITLYLKEKKYSPCAWEVVRAEIMRSFSLSTNLQESLRSKEAKFVAAWTLKAAAGSGSGSDTICIGYHANNSTDTVDTVLEKNVTVTHSVNLLEDSHNGKLCRLKGIAPLQLGKCNIAGWLLGNPECDPLLPVRSWSYIVETPNSENGICYPGDFIDYEELREQLSSVSSFERFEIFPKESSWPNHNTNGVTAACSHEGKSSFYRNLLWLTEKEGSYPKLKNSYVNKKGKEVLVLWGIHHPPNSKEQQNLYQNENAYVSVVTSNYNRRFTPEIAERPKVRDQAGRMNYYWTLLKPGDTIIFEANGNLIAPMYAFALSRGFGSGIITSNASMHECNTKCQTPLGAINSSLPYQNIHPVTIGECPKYVRSAKLRMVTGLRNNPSIQSRFEDKTHTCPPCPAPELLGGPSVFLFPPKPKDQLMISRTPEVTCVVVDVSHEDPEVKFNWYVDGVEVHNAKTKPREEQYNSTYRVVSVLTVLHQDWLNGKEYKCKVSNKALPAPIEKTISKAKGQPREPQVYTLPPSRDELTKNQVSLTCLVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFLYSKLTVDKSRWQQGNVFSCSVLHEALHNHYTQKSLSLSPGK SEQ ID NO: 50: Amino acids of hIFNα-Pan-E7(HPV)-Fc in a homodimer CDLPQTHSLGSRRTLMLLAQMRRISLFSCLKDRHDFGFPQEEFGNQFQKAETIPVLHEMIQQIFNLFSTKDSSAAWDETLLDKFYTELYQQLNDLEACVIQGVGVTETPLMKEDSILAVRKYFQRIT LYLKEKKYSPCAWEVVRAEIMRSFSLSTNLQESLRSKEAKFVAAWTLKAAAGSGSGSMHGDTPTLHEYMLDLQPETTDLYCYEQLNDSSEEEDEIDGPAGQAEPDRAHYNIVTFCCKCDSTLRLCVQS THVDIRTLEDLLMGTLGIVCPICSQKPFEDKTHTCPPCPAPELLGGPSVFLFPPKPKDQLMISRTPEVTCVVVDVSHEDPEVKFNWYVDGVEVHNAKTKPREEQYNSTYRVVSVLTVLHQDWLNGKE YKCKVSNKALAPIEKTISKAKGQPREPQVYTLPPSRDELTKNQVSLTCLVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFLYSKLTVDKSRWQQGNVFSCSVLHEALHNHYTQKSLSLSPGK SEQ ID NO: 51: First chain hIFNα-Fc-hole amino acid sequence in the heterodimer CDLPQTHSLGSRRTLMLLAQMRRISLFSCLKDRHDFGFPQEEFGNQFQKAETIPVLHEMIQQIFNLFSTKDSSAAWDETLLDKFYTELYQQLNDLEACVIQGV GVTETPLMKEDSILAVRKYFQRITLYLKEKKYSPCAWEVVRAEIMRSFSLSTNLQESLRSKESGGGGSGGGGSGGGGSGGGGRTDKTHTCPPCPAPELLGGPSV FLFPPKPKDTLMISRTPEVTCVVVDVSHEDPEVKFNWYVDGVEVHNAKTKPREEQYNSTYRVVSVLTVLHQDWLNGKEYKCKVSNKALPAPIEKTISKAKGQP REPQVCTLPPSRDELTKNQVSLSCAVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLVSKLTVDKSRWQQGNVFSCSVMHEALHNHYTQKSLSLSPGK SEQ ID NO: 52: Amino acid sequence of the second chain Pan-Pres1-Fc-knob in the heterodimeric hIFNα-Pan-Pres1-Fc AKFVAAWTLKAAAGSGSGSMGQNLSTSNPLGFFPDHQLDPAFRANTANPDWDFNPNKDTWPDANKVGAGAFGLGFTPPHGGLLGWSPQAQGILQTLPANPPPASTNRQTGRQPTPLSPPLRNTHPQAFEDKTHTCPPCPAPELLGGPSVFLFPPKPKDTLMISRTPEVTCVVVDVSHE DPEVKFNWYVDGVEVHNAKTKPREEQYNSTYRVVSVLTVLHQDWLNGKEYKCKVSNKALPAPIEKTISKAKGQPREPQVYTLPPCRDELTKNQVSLWCLVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLYSKLTVDKSRWQQGNVFSCSVMHEALHNHYTQKSLSLSPGK SEQ ID NO: 53: Amino acid sequence of the second chain Pan-RBD(SARS-CoV-2)-Fc-knob in the heterodimeric hIFNα-Pan-RBD(SARS-CoV-2)-Fc AKFVAAWTLKAAAGSGSGSRVQPTESIVRFPNITNLCPFGEVFNATRFASVYAWNRKRISNCVADYSVLYNSASFSTFKCYGVSPTKLNDLCFTNVYADSFVIRGDEVRQIAPGQTG KIADYNYKLPDDFTGCVIAWNSNNLDSKVGGNYNYLYRLFRKSNLKPFERDISTEIYQAGSTPCNGVEGFNCYFPLQSYGFQPTNGVGYQPYRVVVLSFELLHAPATVCGPKKSTNLV KNKCVNFFEDKTHTCPPCPAPELLGGPSVFLFPPKPKDTLMISRTPEVTCVVVDVSHEDPEVKFNWYVDGVEVHNAKTKPREEQYNSTYRVVSVLTVLHQDWLNGKEYKCKVSNKALP APIEKTISKAKGQPREPQVYTLPPCRDELTKNQVSLWCLVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLYSKLTVDKSRWQQGNVFSCSVMHEALHNHYTQKSLSLSPGK SEQ ID NO: 54: Amino acid sequence of the second chain Pan-HA-Fc-knob in the heterodimeric hIFNα-Pan-HA-Fc AKFVAAWTLKAAAGSGSGSDTICIGYHANNSTDTVDTVLEKNVTVTHSVNLLEDSHNGKLCRLKGIAPLQLGKCNIAGWLLGNPECDPLLPVRSWSYIVETPNSENGICYPGDFIDYEELREQLSSVSSFERFEIFPKESSWP NHNTNGVTAACSHEGKSSFYRNLLWLTEKEGSYPKLKNSYVNKKGKEVLVLWGIHHPPNSKEQQNLYQNENAYVSVVTSNYNRRFTPEIAERPKVRDQAGRMNYYWTLLKPGDTIIFEANGNLIAPMYAFALSRGGFGSGIITSN ASMHECNTKCQTPLGAINSSLPYQNIHPVTIGECPKYVRSAKLRMVTGLRNNPSIQSRFEDKTHTCPPCPAPELLGGPSVFLFPPKPKDTLMISRTPEVTCVVVDVSHEDPEVKFNWYVDGVEVHNAKTKPREEQYNSTYRVV SVLTVLHQDWLNGKEYKCKVSNKALPAPIEKTISKAKGQPREPQVYTLPPCRDELTKNQVSLWCLVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLYSKLTVDKSRWQQGNVFSCSVMHEALHNHYTQKSLSLSPGK SEQ ID NO: 55: Amino acid sequence of the second chain Pan-HA-Fc-knob in the heterodimeric hIFNα-Pan-E7(HPV)-Fc AKFVAAWTLKAAAGSGSGSMHGDTPTLHEYMLDLQPETTDLYCYEQLNDSSEEEDEIDGPAGQAEPDRAHYNIVTFCCKCDSTLRLCVQSTHVDIRTLEDLLMGTLGIVCPICSQKPFEDKTHTCPPCPAPELLGGPSVFLFPPKPKDTLMISRTPEVTCVVVDVSHEDPEVK FNWYVDGVEVHNAKTKPREEQYNSTYRVVSVLTVLHQDWLNGKEYKCKVSNKALPAPIEKTISKAKGQPREPQVYTLPPCRDELTKNQVSLWCLVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLYSKLTVDKSRWQQGNVFSCSVMHEALHNHYTQKSLSLSPGK 6. Human mutant IFN vaccine hmIFNα-Pan-Antigen-Fc sequence: SEQ ID NO: 56: hmIFNα-Pres1-Fc amino acid sequence in homodimer CDLPQTHSLGSRRTLMLLAQMRRISLFSCLKDRHDFGFPQEEFGNQFQKAETIPVLHEMIQQIFNLFSTKDSSAAWDETLLDKFYTELYQQLNDLEACVIQGVGVTETPLMKEDSILAVRKYFRRITLY LKEKKYSPCAWEVVRAEIMRSFSLSTNLQESLRSKEGGGGSGGGGSGGGGSRTMGQNLSTSNPLGFFPDHQLDPAFRANTANPDWDFNPNKDTWPDANKVGAGAFGLGFTPPHGGLLGWSPQAQGILQTL PANPPPASTNRQTGRQPTPLSPPLRNTHPQAFEDKTHTCPPCPAPELLGGPSVFLFPPKPKDQLMISRTPEVTCVVVDVSHEDPEVKFNWYVDGVEVHNAKTKPREEQYNSTYRVVSVLTVLHQDWLNG KEYKCKVSNKALAPIEKTISKAKGQPREPQVYTLPPSRDELTKNQVSLTCLVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFLYSKLTVDKSRWQQGNVFSCSVLHEALHNHYTQKSLSLSPGK SEQ ID NO: 57: hmIFNα-RBD (SARS-CoV-2)-Fc amino acid sequence in homodimer CDLPQTHSLGSRRTLMLLAQMRRISLFSCLKDRHDFGFPQEEFGNQFQKAETIPVLHEMIQQIFNLFSTKDSSAAWDETLLDKFYTELYQQLNDLEACVIQGVGVTETPMKEDSILAVRKYFRRITLYLKEKKYSPCAWEVVRAEIMRSFSLSTNLQ ESLRSKEGGGGSGGGGSGGGGSRTRVQPTESIVRFPNITNLCPFGEVFNATRFASVYAWNRKRISNCVADYSVLYNSASFSTFKCYGVSPTKLNDLCFTNVYADSFVIRGDEVRQIAPGQTGKIADYNYKLPDDFTGCVIAWNSNNLDSKVGGNYNYL YRLFRKSNLKPFERDISTEIYQAGSTPCNGVEGFNCYFPLQSYGFQPTNGVGYQPYRVVVLSFELLHAPATVCGPKKSTNLVKNKCVNFFEDKTHTCPPCPAPELLGGPSVFLFPPKPKDQLMISRTPEVTCVVVDVSHEDPEVKFNWYVDGVEVHNA KTKPREEQYNSTYRVVSVLTVLHQDWLNGKEYKCKVSNKALPAPIEKTISKAKGQPREPQVYTLPPSRDELTKNQVSLTCLVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFLYSKLTVDKSRWQQGNVFSCSVLHEALHNHYTQKSLSLSPGK SEQ ID NO: 58: hmIFNα-HA-Fc amino acid sequence in homodimer CDLPQTHSLGSRRTLMLLAQMRRISLFSCLKDRHDFGFPQEEFGNQFQKAETIPVLHEMIQQIFNLFSTKDSSAAWDETLLDKFYTELYQQLNDLEACVIQGVGVTETPLMKEDSILAVRKYFRRITLYLKEKKYSPCAWEVVRAEIMRSFSLSTNLQESLRSKEGGGGSGGGGSGGGGSRTDT ICIGYHANNSTDTVDTVLEKNVTVTHSVNLLEDSHNGKLCRLKGIAPLQLGKCNIAGWLLGNPECDPLLPVRSWSYIVETPNSENGICYPGDFIDYEELREQLSSVSSFERFEIFPKESSWPNHNTNGVTAACSHEGKSSFYRNLLWLTEKEGSYPKLKNSYVNKKGKEVLVLWGIHHPPNSKE QQNLYQNENAYVSVVTSNYNRRFTPEIAERPKVRDQAGRMNYYWTLLKPGDTIIFEANGNLIAPMYAFALSRGFGSGIITSNASMHECNTKCQTPLGAINSSLPYQNIHPVTIGECPKYVRSAKLRMVTGLRNNPSIQSRFEDKTHTCPPCPAPELLGGPSVFLFPPKKDQLMISRTPEVTCV VVDVSHEDPEVKFNWYVDGVEVHNAKTKPREEQYNSTYRVVSVLTVLHQDWLNGKEYKCKVSNKALPAPIEKTISKAKGQPREPQVYTLPPS RDELTKNQVSLTCLVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFLYSKLTVDKSRWQQGNVFSCSVLHEALHNHYTQKSLSLSPGK SEQ ID NO: 59: hmIFNα-E7(HPV)-Fc amino acid sequence in homodimer CDLPQTHSLGSRRTLMLLAQMRRISLFSCLKDRHDFGFPQEEFGNQFQKAETIPVLHEMIQQIFNLFSTKDSSAAWDETLLDKFYTELYQQLNDLEACVIQGVGVTETPLMKEDSILAVRKYFRRITLYLKEKKYSPCAWEVVRAEIMRSFSLSTNLQESLRSKEGGGGSGGGGSGGGGSRTMHGDTPTLHEYMLDLQPETTDLYCYEQLNDSSEEEDEIDGPAGQAEPDRAHYNIVTFCCKCDSTLRLCVQSTHVDIRTLEDLLMGTLGIVCPICSQKPFEDKTHTCPPCPAPELLGGPSVFLFPPKPKDQLMISRTPEVTCVVVDVSHEDPEVKFNWYVDGVEVHNAKTKPREEQYNSTYRVVSVLTVLHQDWLNGKEYKCKVSNKALPAPIEKTISKAKGQPREPQVYTLPPSRDELTKNQVSLTCLVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFLYSKLTVDKSRWQQGNVFSCSVLHEALHNHYTQKSLSLSPGK SEQ ID NO: 60: Amino acid sequence of the first chain hmIFN-Fc-hole in a heterodimer CDLPQTHSLGSRRTLMLLAQMRRISLFSCLKDRHDFGFPQEEFGNQFQKAETIPVLHEMIQQIFNLFSTKDSSAAWDETLLDKFYTELYQQLNDLEACVIQGVGVTETPLMKEDSILAVRKYFRRITLYLKEKKYSPCAWEVVRAEIMRSFSLSTNLQESLRSKESGGGGSGGGGSGGGGSGGGGRTDKTHTCPPCPAPELLGGPSVFLFPPKPKDTLMISRTPEVTCVVVDVSHEDPEVKFNWYVDGVEVHNAKTKPREEQYNSTYRVVSVLTVLHQDWLNGKEYKCKVSNKALPAPIEKTISKAKGQPREPQVCTLPPSRDELTKNQVSLSCAVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLVSKLTVDKSRWQQGNVFSCSVMHEALHNHYTQKSLSLSPGK SEQ ID NO: 61: Amino acid sequence of the second chain Pres1-Fc-knob in the heterodimer hmIFNα-Pres1-Fc MGQNLSTSNPLGFFPDHQLDPAFRANTANPDWDFNPNKDTWPDANKVGAGAFGLGFTPPHGGLLGWSPQAQGILQTLPANPPPASTNRQTGRQPTPLSPPLRNTHPQAFEDKTHTCPPCPAPELLGGPSVFLFPPKPKDTLMISRTPEVTCVVVDVSHEDPEVKFNWY VDGVEVHNAKTKPREEQYNSTYRVVSVLTVLHQDWLNGKEYKCKVSNKALPAPIEKTISKAKGQPREPQVYTLPPCRDELTKNQVSLWCLVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLYSKLTVDKSRWQQGNVFSCSVMHEALHNHYTQKSLSLSPGK SEQ ID NO: 62: Second chain RBD(SARS-CoV-2)-Fc-knob amino acid sequence in heterodimeric hmIFNα-RBD(SARS-CoV-2)-Fc RVQPTESIVRFPNITNLCPFGEVFNATRFASVYAWNRKRISNCVADYSVLYNSASFSTFKCYGVSPTKLNDLCFTNVYADSFVIRGDEVRQIAPGQTGKIADYNYKLPDDFTG CVIAWNSNNLDSKVGGNYNYLYRLFRKSNLKPFERDISTEIYQAGSTPCNGVEGFNCYFPLQSYGFQPTNGVGYQPYRVVVLSFELLHAPATVCGPKKSTNLVKNKCVNFFED KTHTCPPCPAPELLGGPSVFLFPPKPKDTLMISRTPEVTCVVVDVSHEDPEVKFNWYVDGVEVHNAKTKPREEQYNSTYRVVSVLTVLHQDWLNGKEYKCKVSNKALPAPIEK TISKAKGQPREPQVYTLPPCRDELTKNQVSLWCLVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLYSKLTVDKSRWQQGNVFSCSVMHEALHNHYTQKSLSLSPGK SEQ ID NO: 63: Amino acid sequence of the second chain HA-Fc-knob in the heterodimeric hmIFNα-HA-Fc DTICIGYHANNSTDTVDTVLEKNVTVTHSVNLLEDSHNGKLCRLKGIAPLQLGKCNIAGWLLGNPECDPLLPVRSWSYIVETPNSENGICYPGDFIDYEELREQLSSVSSFERFEIFPKESSWPNHNTNGVTAACSHE GKSSFYRNLLWLTEKEGSYPKLKNSYVNKKGKEVLVLWGIHPPNSKEQQNLYQNENAYVSVVTSNYNRRFTPEIAERPKVRDQAGRMNYYWTLLKPGDTIIFEANGNLIAPMYAFALSRGGFGSGIITSNASMHECNTK CQTPLGAINSSLPYQNIHPVTIGECPKYVRSAKLRMVTGLRNNPSIQSRFEDKTHTCPPCPAPELLGGPSVFLFPPKPKDTLMISRTPEVTCVVDVSHEDPEVKFNWYVDGVEVHNAKTKPREEQYNSTYRVVSVLTV LHQDWLNGKEYKCKVSNKALPAPIEKTISKAKGQPREPQVYTLPPCRDELTKNQVSLWCLVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLYSKLTVDKSRWQQGNVFSCSVMHEALHNHYTQKSLSLSPGK SEQ ID NO: 64: Amino acid sequence of the second chain HA-Fc-knob in the heterodimeric hmIFNα-E7(HPV)-Fc MHGDTPTLHEYMLDLQPETTDLYCYEQLNDSSEEEDEIDGPAGQAEPDRAHYNIVTFCCKCDSTLRLCVQSTHVDIRTLEDLLMGTLGIVCPICSQKPFEDKTHTCPPCPAPELLGGPSVFLFPPKPKDTLMISRTPEVTCVVDVSHEDPEVKFNWYVDGVE VHNAKTKPREEQYNSTYRVVSVLTVLHQDWLNGKEYKCKVSNKALPAPIEKTISKAKGQPREPQVYTLPPCRDELTKNQVSLWCLVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLYSKLTVDKSRWQQGNVFSCSVMHEALHNHYTQKSLSLSPGK 7. Vaccine containing Pan epitopes of human mutant IFNs hmIFNα-Pan epitope-antigen-Fc sequence SEQ ID NO: 65: hmIFNα-Pan-Pres1-Fc amino acid sequence in homodimer CDLPQTHSLGSRRTLMLLAQMRRISLFSCLKDRHDFGFPQEEFGNQFQKAETIPVLHEMIQQIFNLFSTKDSSAAWDETLLDKFYTELYQQLNDLEACVIQGVGVTETPLMKEDSILAVRKYFRRITLYL KEKKYSPCAWEVVRAEIMRSFSLSTNLQESLRSKEAKFVAAWTLKAAAGSGSGSMGQNLSTSNPLGFFPDHQLDPAFRANTANPDWDFNPNKDTWPDANKVGAGAFGLGFTPPHGGLLGWSPQAQGILQT LPANPPPASTNRQTGRQPTPLSPPLRNTHPQAFEDKTHTCPPCPAPELLGGPSVFLFPPKPKDQLMISRTPEVTCVVVDVSHEDPEVKFNWYVDGVEVHNAKTKPREEQYNSTYRVVSVLTVLHQDWLNG KEYKCKVSNKALAPIEKTISKAKGQPREPQVYTLPPSRDELTKNQVSLTCLVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFLYSKLTVDKSRWQQGNVFSCSVLHEALHNHYTQKSLSLSPGK SEQ ID NO: 66: hmIFNα-Pan-RBD(SARS-CoV-2)-Fc amino acid sequence in homodimer CDLPQTHSLGSRRTLMLLAQMRRISLFSCLKDRHDFGFPQEEFGNQFQKAETIPVLHEMIQQIFNLFSTKDSSAAWDETLLDKFYTELYQQLNDLEACVIQGVGVTETPMKEDSILAVRKYFRRITLYLKEKKYSPCAWEVVRAEIMRSFSLSTNLQ ESLRSKEAKFVAAWTLKAAAGSGSGSRVQPTESIVRFPNITNLCPFGEVFNATRFASVYAWNRKRISNCVADYSVLYNSASFSTFKCYGVSPTKLNDLCFTNVYADSFVIRGDEVRQIAPGQTGKIADYNYKLPDDFTGCVIAWNSNNLDSKVGGNYNY LYRLFRKSNLKPFERDISTEIYQAGSTPCNGVEGFNCYFPLQSYGFQPTNGVGYQPYRVVVLSFELLHAPATVCGPKKSTNLVKNKCVNFFEDKTHTCPPCPAPELLGGPSVFLFPPKPKDQLMISRTPEVTCVVDVSHEDPEVKFNWYVDGVEVHNA KTKPREEQYNSTYRVVSVLTVLHQDWLNGKEYKCKVSNKALPAPIEKTISKAKGQPREPQVYTLPPSRDELTKNQVSLTCLVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFLYSKLTVDKSRWQQGNVFSCSVLHEALHNHYTQKSLSLSPGK SEQ ID NO: 67: hmIFNα-Pan-HA-Fc amino acid sequence in homodimer CDLPQTHSLGSRRTLMLLAQMRRISLFSCLKDRHDFGFPQEEFGNQFQKAETIPVLHEMIQQIFNLFSTKDSSAAWDETLLDKFYTELYQQLNDLEACVIQGVGVTETPLMKEDSILAVRKYFRRITLYLKEKKYSPCAWEVVRAEIMRSFSLSTNLQESLRSKEAKFVAAWTLKAAAGSGSGS DTICIGYHANNSTDTVDTVLEKNVTVTHSVNLLEDSHNGKLCRLKGIAPLQLGKCNIAGWLLGNPECDPLLPVRSWSYIVETPNSENGICYPGDFIDYEELREQLSSVSSFERFEIFPKESSWPNHNTNGVTAACSHEGKSSFYRNLLWLTEKEGSYPKLKNSYVNKKGKEVLVLWGIHHPPNSK EQQNLYQNENAYVSVVTSNYNRRFTPEIAERPKVRDQAGRMNYYWTLLKPGDTIIFEANGNLIAPMYAFALSRGFGSGIITSNASMHECNTKCQTPLGAINSSLPYQNIHPVTIGECPKYVRSAKLRMVTGLRNNPSIQSRFEDKTHTCPPCPAPELLGGPSVFLFPPKPKDQLMISRTPEVTC VVVDVSHEDPEVKFNWYVDGVEVHNAKTKPREEQYNSTYRVVSVLTVLHQDWLNGKEYKCKVSNKALPAPIEKTISKAKGQPREPQVYTLPP SRDELTKNQVSLTCLVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFLYSKLTVDKSRWQQGNVFSCSVLHEALHNHYTQKSLSLSPGK SEQ ID NO: 68: hmIFNα-Pan-E7(HPV)-Fc amino acid sequence in homodimer CDLPQTHSLGSRRTLMLLAQMRRISLFSCLKDRHDFGFPQEEFGNQFQKAETIPVLHEMIQQIFNLFSTKDSSAAWDETLLDKFYTELYQQLNDLEACVIQGVGVTETPLMKEDSILAVRKYFRRIT LYLKEKKYSPCAWEVVRAEIMRSFSLSTNLQESLRSKEAKFVAAWTLKAAAGSGSGSMHGDTPTLHEYMLDLQPETTDLYCYEQLNDSSEEEDEIDGPAGQAEPDRAHYNIVTFCCKCDSTLRLCVQS THVDIRTLEDLLMGTLGIVCPICSQKPFEDKTHTCPPCPAPELLGGPSVFLFPPKPKDQLMISRTPEVTCVVVDVSHEDPEVKFNWYVDGVEVHNAKTKPREEQYNSTYRVVSVLTVLHQDWLNGKE YKCKVSNKALAPIEKTISKAKGQPREPQVYTLPPSRDELTKNQVSLTCLVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFLYSKLTVDKSRWQQGNVFSCSVLHEALHNHYTQKSLSLSPGK SEQ ID NO: 69: First chain hmIFNα4-Fc-hole amino acid sequence in the heterodimer CDLPQTHSLGSRRTLMLLAQMRRISLFSCLKDRHDFGFPQEEFGNQFQKAETIPVLHEMIQQIFNLFSTKDSSAAWDETLLDKFYTELYQQLNDLEACVIQGV GVTETPLMKEDSILAVRKYFRRITLYLKEKKYSPCAWEVVRAEIMRSFSLSTNLQESLRSKESGGGGSGGGGSGGGGSGGGGRTDKTHTCPPCPAPELLGGPSV FLFPPKPKDTLMISRTPEVTCVVVDVSHEDPEVKFNWYVDGVEVHNAKTKPREEQYNSTYRVVSVLTVLHQDWLNGKEYKCKVSNKALPAPIEKTISKAKGQP REPQVCTLPPSRDELTKNQVSLSCAVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLVSKLTVDKSRWQQGNVFSCSVMHEALHNHYTQKSLSLSPGK SEQ ID NO: 70: Amino acid sequence of the second chain Pan-Pres1-Fc-knob in the heterodimer hmIFNα-Pan-Pres1-Fc AKFVAAWTLKAAAGSGSGSMGQNLSTSNPLGFFPDHQLDPAFRANTANPDWDFNPNKDTWPDANKVGAGAFGLGFTPPHGGLLGWSPQAQGILQTLPANPPPASTNRQTGRQPTPLSPPLRNTHHPQAFEDKTHTCPPCPAPELLGGPSVFLFPPKPKDTLMISRTPEVTCVVVDVSHE DPEVKFNWYVDGVEVHNAKTKPREEQYNSTYRVVSVLTVLHQDWLNGKEYKCKVSNKALPAPIEKTISKAKGQPREPQVYTLPPCRDELTKNQVSLWCLVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLYSKLTVDKSRWQQGNVFSCSVMHEALHNHYTQKSLSLSPGK SEQ ID NO: 71: Amino acid sequence of the second chain Pan-RBD(SARS-CoV-2)-Fc-knob in the heterodimeric hmIFNα-Pan-RBD(SARS-CoV-2)-Fc AKFVAAWTLKAAAGSGSGSRVQPTESIVRFPNITNLCPFGEVFNATRFASVYAWNRKRISNCVADYSVLYNSASFSTFKCYGVSPTKLNDLCFTNVYADSFVIRGDEVRQIAPGQTG KIADYNYKLPDDFTGCVIAWNSNNLDSKVGGNYNYLYRLFRKSNLKPFERDISTEIYQAGSTPCNGVEGFNCYFPLQSYGFQPTNGVGYQPYRVVVLSFELLHAPATVCGPKKSTNLV KNKCVNFFEDKTHTCPPCPAPELLGGPSVFLFPPKPKDTLMISRTPEVTCVVVDVSHEDPEVKFNWYVDGVEVHNAKTKPREEQYNSTYRVVSVLTVLHQDWLNGKEYKCKVSNKALP APIEKTISKAKGQPREPQVYTLPPCRDELTKNQVSLWCLVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLYSKLTVDKSRWQQGNVFSCSVMHEALHNHYTQKSLSLSPGK SEQ ID NO: 72: Amino acid sequence of the second chain Pan-HA-Fc-knob in the heterodimer hmIFNα-Pan-HA-Fc AKFVAAWTLKAAAGSGSGSDTICIGYHANNSTDTVDTVLEKNVTVTHSVNLLEDSHNGKLCRLKGIAPLQLGKCNIAGWLLGNPECDPLLPVRSWSYIVETPNSENGICYPGDFIDYEELREQLSSVSSFERFEIFPKESSWP NHNTNGVTAACSHEGKSSFYRNLLWLTEKEGSYPKLKNSYVNKKGKEVLVLWGIHPPNSKEQQNLYQNENAYVSVVTSNYNRRFTPEIAERPKVRDQAGRMNYYWTLLKPGDTIIFEANGNLIAPMYAFALSRGGFGSGIITSN ASMHECNTKCQTPLGAINSSLPYQNIHPVTIGECPKYVRSAKLRMVTGLRNNPSIQSRFEDKTHTCPPCPAPELLGGPSVFLFPPKPKDTLMISRTPEVTCVVVDVSHEDPEVKFNWYVDGVEVHNAKTKPREEQYNSTYRVV SVLTVLHQDWLNGKEYKCKVSNKALPAPIEKTISKAKGQPREPQVYTLPPCRDELTKNQVSLWCLVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLYSKLTVDKSRWQQGNVFSCSVMHEALHNHYTQKSLSLSPGK SEQ ID NO: 73: Amino acid sequence of the second chain Pan-HA-Fc-knob in the heterodimeric hmIFNα-Pan-E7(HPV)-Fc AKFVAAWTLKAAAGSGSGSMHGDTPTLHEYMLDLQPETTDLYCYEQLNDSSEEEDEIDGPAGQAEPDRAHYNIVTFCCKCDSTLRLCVQSTHVDIRTLEDLLMGTLGIVCPICSQKPFEDKTHTCPPCPAPELLGGPSVFLFPPKPKDTLMISRTPEVTCVVVDVSHEDPEVK FNWYVDGVEVHNAKTKPREEQYNSTYRVVSVLTVLHQDWLNGKEYKCKVSNKALPAPIEKTISKAKGQPREPQVYTLPPCRDELTKNQVSLWCLVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLYSKLTVDKSRWQQGNVFSCSVMHEALHNHYTQKSLSLSPGK 8. Antibody sequences instead of Fc SEQ ID NO: 20: ScFv(PD-L1) amino acid sequence DIQMTQSPSSLSASVGDRVTITCRASQDVSTAVAWYQQKPGKAPKLLIYSASFLYSGVPSRFSGSGSGTDFTLTISSLQPEDFATYYCQQYLYHPATFGQGTKVEIKRGGGGSGGGSGG GGSEVQLVESGGGLVQPGGSLRLSCAASGFTFSDSWIHWVRQAPGKGLEWVAWISPYGGSTYYADSVKGRFTISADTSKNTAYLQMNSLRAEDTAVYYCARRHWPGGFDYWGQGTLVTVSA SEQ ID NO: 74: Anti-PD-L1 VH amino acid sequence EVQLVESGGGLVQPGGSLRLSCAASGFTFSDSWIHWVRQAPGKGLEWVAWISPYGGSTYYADSVKGRFTISADTSKNTAYLQMNSLRAEDTAVYYCARRHWPGGFDYWGQGTLVTVSA SEQ ID NO: 75:Anti-PD-L1 VL amino acid sequence DIQMTQSPSSLSASVGDRVTITCRASQDVSTAVAWYQQKPGKAPKLLIYSASFLYSGVPSRFSGSGSGTDFTLTISSLQPEDFATYYCQQYLYHPATFGQGTKVEIKR 9. Other viral antigen sequences SEQ ID NO: 76: Amino acid sequence of SARS-CoV-2 Spike protein VNLTTRTQLPPAYTNSFTRGVYYPDKVFRSSVLHSTQDLFLPFFSNVTWFHAIHVSGTNGTKRFDNPVLPFNDGVYFASTEKSNIIRGWIFGTTLDSKTQSLLIVNNATNVVIKVCEFQFCNDPFLGVYYHKNNKSWMESEFRVYSSANNCTFEYVSQPFLMDLEGKQGNFKNLREFVFKNIDGYFKIYSKHTPINLVRDLPQGFSALEPLVDLPIGINITRFQTLLALHRSYLTPGDSSSGWTAGAAAYYVGYLQPRTFLLKYNENGTITDAVDCALDPLSETKCTLKSFTVEKGIYQTSNFRVQPTESIVRFPNITNLCPFGEVFNATRFASVYAWNRKRISNCVADYSVLYNSASFSTFKCYGVSPTKLNDLCFTNVYADSFVIRGDEVRQIAPGQTGKIADYNYKLPDDFTGCVIAWNSNNLDSKVGGNYNYLYRLFRKSNLKPFERDISTEIYQAGSTPCNGVEGFNCYFPLQSYGFQPTNGVGYQPYRVVVLSFELLHAPATVCGPKKSTNLVKNKCVNFNFNGLTGTGVLTESNKKFLPFQQFGRDIADTTDAVRDPQTLEILDITPCSFGGVSVITPGTNTSNQVAVLYQDVNCTEVPVAIHADQLTPTWRVYSTGSNVFQTRAGCLIGAEHVNNSYECDIPIGAGICASYQTQTNSPRRARSVASQSIIAYTMSLGAENSVAYSNNSIAIPTNFTISVTTEILPVSMTKTSVDCTMYICGDSTECSNLLLQYGSFCTQLNRALTGIAVEQDKNTQEVFAQVKQIYKTPPIKDFGGFNFSQILPDPSKPSKRSFIEDLLFNKVTLADAGFIKQYGDCLGDIAARDLICAQKFNGLTVLPPLLTDEMIAQYTSALLAGTITSGWTFGAGAALQIPFAMQMAYRFNGIGVTQNVLYENQKLIANQFNSAIGKIQDSLSSTASALGKLQD VVNQNAQALNTLVKQLSSNFGAISSVLNDILSRLDKVEAEVQIDRLITGRLQSLQTYVTQQLIRAAEIRASANLAATKMSECVLGQSKRVDFCGKGYHLMSFPQSAPHGVVFLHVTYVPAQEKNFTTAPAICHDGKAHFPREGVFVSNGTHWFVTQRNFYE PQIITTDNTFVSGNCDVVIGIVNNTVYDPLQPELDSFKEELDKYFKNHTSPDVDLGDISGINASVVNIQKEIDRLNEVAKNLNESLIDLQELGKYEQYIKWPWYIWLGFIAGLIAIVMVTIMLCCMTSCCSCLKGCCSCGSCCKFDEDDSEPVLKGVKLHYT SEQ ID NO: 77: Amino acid sequence of SARS-CoV-2 S1 protein VNLTTRTQLPPAYTNSFTRGVYYPDKVFRSSVLHSTQDLFLPFFSNVTWFHAIHVSGTNGTKRFDNPVLPFNDGVYFASTEKSNIIRGWIFGTTLDSKTQSLLIVNNATNVVIKVCEFQFCNDPFLGVYYHKNNKSWMESEFRVYSSANNCTFEYVSQPFLMDLEGK QGNFKNLREFVFKNIDGYFKIYSKHTPINLVRDLPQGFSALEPLVDLPIGINITRFQTLLALHRSYLTPGDSSSGWTAGAAAYYVGYLQPRTFLLKYNENGTITDAVDCALDPLSETKCTLKSFTVEKGIYQTSNFRVQPTESIVRFPNITNLCPFGEVFNATRFASV YAWNRKRISNCVADYSVLYNSASFSTFKCYGVSPTKLNDLCFTNVYADSFVIRGDEVRQIAPGQTGKIADYNYKLPDDFTGCVIAWNSNLDSKVGGNYNYLYRLFRKSNLKPFERDISTEIYQAGSTPCNGVEGFNCYFPLQSYGFQPTNGVGYQPYRVVVLSFEL LHAPATVCGPKKSTNLVKNKCVNFNFGLTGTGVLTESNKKFLPFQQFGRDIADTTDAVRDPQTLEILDITPCSFGGVSVITPGTNTSNQVAVLYQDVNCTEVPVAIHADQLTTPWRVYSTGSNVFQTRAGCLIGAEHVNNSYECDIPIGAGICASYQTQTNSPRRAR SEQ ID NO: 78:SARS-CoV-2 original amino acid sequence of RBD protein RVQPTESIVRFPNITNLCPFGEVFNATRFASVYAWNRKRISNCVADYSVLYNSASFSTFKCYGVSPTKLNDLCFTNVYADSFVIRGDEVRQIAPGQTGKIADYNYKLPDDFTGCVIAWNSNNLDSKVGGNYLYRLFRKSNLKPFERDISTEIYQAGSTPCNGVEGFNCYFPLQSYGFQPTNGVGYQPYRVVVLSFELLHAPATVCGPKKSTNLVKNKCVNF SEQ ID NO: 79: Amino acid sequence of the SARS-CoV-2 UK variant (B.1.1.7, Alpha) RBD protein RVQPTESIVRFPNITNLCPFGEVFNATRFASVYAWNRKRISNCVADYSVLYNSASFSTFKCYGVSPTKLNDLCFTNVYADSFVIRGDEVRQIAPGQTGKIADYNYKLPDDF TGCVIAWNSNNLDSKVGGNYNYLYRLFRKSNLKPFERDISTEIYQAGSTPCNGVEGFNCYFPLQSYGFQPTYGVGYQPYRVVVLSFELLHAPATVCGPKKSTNLVKNKCVNF SEQ ID NO: 80: Amino acid sequence of the RBD protein of SARS-CoV-2 South African variant (B.1.351, Beta) RVQPTESIVRFPNITNLCPFGEVFNATRFASVYAWNRKRISNCVADYSVLYNSASFSTFKCYGVSPTKLNDLCFTNVYADSFVIRGDEVRQIAPGQTGNIADYNYKLPDDF TGCVIAWNSNNLDSKVGGNYNYLYRLFRKSNLKPFERDISTEIYQAGSTPCNGVKGFNCYFPLQSYGFQPTYGVGYQPYRVVVLSFELLHAPATVCGPKKSTNLVKNKCVNF SEQ ID NO: 81: SARS-CoV-2 Brazilian variant (P.1) RBD protein amino acid sequence RVQPTESIVRFPNITNLCPFGEVFNATRFASVYAWNRKRISNCVADYSVLYNSASFSTFKCYGVSPTKLNDLCFTNVYADSFVIRGDEVRQIAPGQTGTIADYNYKLPDDF TGCVIAWNSNNLDSKVGGNYNYLYRLFRKSNLKPFERDISTEIYQAGSTPCNGVKGFNCYFPLQSYGFQPTYGVGYQPYRVVVLSFELLHAPATVCGPKKSTNLVKNKCVNF SEQ ID NO: 82: Amino acid sequence of the RBD protein of the SRAS-CoV-2 California variant (B.1.429) RVQPTESIVRFPNITNLCPFGEVFNATRFASVYAWNRKRISNCVADYSVLYNSASFSTFKCYGVSPTKLNDLCFTNVYADSFVIRGDEVRQIAPGQTGKIADYNYKLPDDF TGCVIAWNSNNLDSKVGGNYNYRYRLFRKSNLKPFERDISTEIYQAGSTPCNGVEGFNCYFPLQSYGFQPTNGVGYQPYRVVVLSFELLHAPATVCGPKKSTNLVKNKCVNF SEQ ID NO: 83: Amino acid sequence of RBD protein of SRAS-CoV-2 Indian B.1.617, B.1.617.1 (Kappa), and B.1.617.3 variants RVQPTESIVRFPNITNLCPFGEVFNATRFASVYAWNRKRISNCVADYSVLYNSASFSTFKCYGVSPTKLNDLCFTNVYADSFVIRGDEVRQIAPGQTGKIADYNYKLPDDF TGCVIAWNSNNLDSKVGGNYNYRYRLFRKSNLKPFERDISTEIYQAGSTPCNGVQGFNCYFPLQSYGFQPTNGVGYQPYRVVVLSFELLHAPATVCGPKKSTNLVKNKCVNF SEQ ID NO: 84: Amino acid sequence of the RBD protein of SRAS-CoV-2 Indian B.1.617.2 (Delta) variant RVQPTESIVRFPNITNLCPFGEVFNATRFASVYAWNRKRISNCVADYSVLYNSASFSTFKCYGVSPTKLNDLCFTNVYADSFVIRGDEVRQIAPGQTGKIADYNYKLPDDF TGCVIAWNSNNLDSKVGGNYNYRYRLFRKSNLKPFERDISTEIYQAGSKPCNGVEGFNCYFPLQSYGFQPTNGVGYQPYRVVVLSFELLHAPATVCGPKKSTNLVKNKCVNF 10. Other tumor antigen sequences: Amino acid sequences of murine Her2 extracellular domains II, III and IV according to the embodiment: SEQ ID NO: 85:Mouse Her2-extracellular domain 2: SRACPPCAPACKDNHCWGESPEDCQILTGTICTSGCARCKGRLPTDCCHEQCAAGCTGPKHSDCLACLHFNHSGICELHCPALVTYNTDTFESMHNPEGRYTFGASCVTTCPYNYLSTEVGSCTLVCPPPNNQEVTAEDGTQRCEKCSKPC SEQ ID NO: 86:Mouse Her2-extracellular domain 3: GCKKIFGSLAFLPESFDGDPSSGIAPLRPEQLQVFETLEEITGYLYISAWPDSLRDLSVFQNLRIIRGRILHDGAYSLTLQGLGIHSLGLRSLRELGSGLALIHRNAHLCFVHTVPWDQLF SEQ ID NO: 87:Mouse Her2-extracellular domain 4: VCNSLCAHGHCWGPGPTQCVNCSHFLRGQECVEECRVWKGLPREYVSDKRCLPCHPECQPQNSSETCFGSEADQCAACAHYKDSSSCVARCPSGVKPDLSYMPIWKYPDEEGICQPCPINCTHSCVDLDERGCP Amino acid sequences of human Her2 extracellular domains II, III and IV according to the embodiment: SEQ ID NO: 88:Human Her2-extracellular domain 2: SRACHPCSPMCKGSRCWGESSEDCQSLTRTVCAGGCARCKGPLPTDCCHEQCAAGCTGPKHSDCLACLHFNHSGICELHCPALVTYNTDTFESMPNPEGRYTFGASCVTACPYNYLSTDVGSCTLVCPLHNQEVTAEDGTQRCEKCSKPC SEQ ID NO: 89:Human Her2-extracellular domain 3: GCKKIFGSLAFLPESFDGDPASNTAPLQPEQLQVFETLEEITGYLYISAWPDSLPDLSVFQNLQVIRGRILHNGAYSLTLQGLGISWLGLRSLRELGSGLALIHHNTHLCFVHTVPW SEQ ID NO: 90: Human Her2 - extracellular domain 4: CHQLCARGHCWGPGPTQCVNCSQFLRGQECVEECRVLQGLPREYVNARHCLPCHPECQPQNGSVTCFGPEADQCVACAHYKDPPFCVARCPSGVKPDLSYMPIWKFPDEEGACQPCPINCTHSCVDLDDKGCP 11. Herpes virus antigen sequence according to the example: SEQ ID NO: 91: VZV Envelope glycoprotein E (aa 31 - 538) SVLRYDDFHTDEDKLDTNSVYEPYYHSDHAESSWVNRGESSRKAYDHNSPYIWPRNDYDGFLENAHEHHGVYNQGRGIDSGERLMQPTQMSAQEDLGDDTGIHVIPTLNGDDRHKIVNVDQRQYGDVFKGDLNPKPQGQRLIEVSVEENHPFTLRAPIQRIYGVRYTETWSFLPSLTCTGDAAPAIQHICLKHTTCFQDVVVDVDCAENTKEDQLAEISYRFQGKKEADQPWIVVNTSTLFDELELDPPEIEPGVLKVLRTEKQYLGVYIWNMRGSDGTSTYATFLVTWKGDEKTRNPTPAVTPQPRGAEFHMWNYHSHVFSVGDTFSLAMHLQYKIHEAPFDLLLEWLYVPIDPTCQPMRLYSTCLYHPNAPQCLSHMNSGCTFTSPHLAQRVASTVYQNCEHADNYTAYCLGISHMEPSFGLILHDGGTTLKFVDTPESLSGLYVFVVYFNGHVEAVAYTVVSTVDHFVNAIEERGFPPTAGQPPATTKPKEITPVNPGTSPLLRY SEQ ID NO: 92: EBV Envelope glycoprotein GP350 (aa 1-425) MEAALLVCQYTIQSLIHLTGEDPGFFNVEIPEFPFYPTCNVCTADVNVTINFDVGGKKHQLDLDFGQLTPHTKAVYQPRGAFGGSENATNLFLLELLGAGELALTMRSKKLPINVTTGEEQQVSLESVDVYFQDVFGTMWCHHAEMQNPVYLIPETVPYIKWDNCNSTNITAVVRAQGLDVTLPLSLPTSAQDSNFSVKTQMLGNEIDIECIMEDGEISQVLPGDNKFNITCSGYESHVPSGGILTSTSPVVTPIPGTGYAYSLRLTPRPVSRFLGNNSILYVFYSGNGPKASGGDYCIQSNIVFSDEIPASQDMPTNTTDITYVGDNATYSVPMVTSEDANSPNVTVTAFWAWPNNTETDFKCKWTLTSGTPSGCENISGAFASNRTFDITVSGLGTAPKTLIITRTATNATTTTHKVIFSKAP SEQ ID NO: 93: HSV-2 Envelope glycoprotein gD (aa 26-339) KYALADPSLKMADPNRFRGKNLPVLDRLTDPPGVKRVYHIQPSLEDPFQPPSIPITVYYAVLERACRSVLLHAPSEAPQIVRGASDEARKHTYNLTIAWYRMGDNCAIPITVMEYTECPYNKSLGVCPIRTQPRWSYYDSFSAVSEDNLGFLMHAPAFETAGTYLRLVKINDWTEITQFILEHRARASCKYALPLRIPPAACLTSKAYQQGVTVDSIGMLPRFIPENQRTVALYSLKIAGWHGPKPPYTSTLLPPELSDTTNATQPELVPEDPEDSALLEDPAGTVSSQIPPNWHIPSIQDVAPHHAPAAPSNP

Brief Description of the Drawings

[0045] [Figure 1]Figure 1. Schematic diagram of the vaccine platform combining the following order in the form of a homodimer: interferon-linked fragment 1-target antigen-immunoglobulin Fc (or antibody); [Diagram 2] Figure 2. Schematic diagram of the vaccine platform combining interferon-linked fragment 1-IgG1-hole, target antigen-IgG1-knob (or antibody) in the form of heterodimers; [Diagram 3] Figure 3. Schematic diagram of the vaccine platform, combining interferon-linked fragment 1-IgG1-knob and target antigen-IgG1-hole (or antibody) in the form of heterodimers, respectively; [Figure 4] Figure 4. Schematic diagram of the vaccine platform, combining the following order in the form of a homodimer: interferon-linked fragment 1-Th cell helper epitope-linked fragment 2-target antigen-immunoglobulin Fc (or antibody); [Diagram 5] Figure 5. Schematic diagram of the vaccine platform combined in the form of heterodimers in the order of interferon-linked fragment 1-IgG1-hole, and Th cell helper epitope-linked fragment 2-target antigen-IgG1-knob (or antibody), respectively; [Figure 6] Figure 6. Schematic diagram of the vaccine platform, in which interferon-linked fragment 1-IgG1-knob and Th cell helper epitope-linked fragment 2-target antigen-IgG1-hole (or antibody) are combined in the form of heterodimers; [Figure 7] Figure 7. Identification of Pres1-Fc and IFN-Pres1-Fc non-denatured proteins by SDS-PAGE electrophoresis. [Figure 8]Figure 8. Compared with free preS1, the fusion proteins preS1-Fc and IFN-preS1-Fc can significantly enhance the immunogenicity of antigen molecules and induce the production of broadly neutralizing antibodies. (a) C57 / BL6 (n=8 / group) mice were inoculated with free Hepatitis B Pres1, Pres1-Fc, and IFNα-Pres1-Fc proteins by subcutaneous immunization, and the level of Pres1-specific antibodies in serum was detected by Elisa at the indicated time. (b) Three HBV genotype stable carrier mice (n=4) were intravenously injected with serum from IFNα-Pres1-Fc protein-immunized mice, and the change of Pres1 antigen in serum was detected 12 hours later. [Figure 9] Figure 9. IFNα-Pres1-Fc can be used as a preventive vaccine for hepatitis B. C57 / BL6 mice were inoculated with free hepatitis B Pres1, Pres1-Fc, and IFNα-Pres1-Fc proteins by subcutaneous immunization, and then infected with 1x1011μg of AAV-HBV1.3 virus by tail vein injection 28 days after inoculation. (a) Serum Anti-Pres1 levels before and 1, 2, 3, and 4 weeks after virus inoculation. (b) Pres1 levels in serum were detected at the indicated time points. (c) Serum HBsAg levels were detected by ELISA at 1, 2, 3, and 4 weeks. (d) Percentage of HBsAg-positive mice after AAV-HBV1.3 virus inoculation. [Figure 10] Figure 10. IFNα-Pres1-Fc was used as a preventive vaccine for chronic type B infection. C57 / BL6 mice were infected with 1x1011μg of AAV-HBV1.3 virus via tail vein injection, and stably infected mice (n=8 / group) were selected 6 weeks after infection and immunized subcutaneously with recombinant Pres1 and IFNα-Pres1-Fc proteins once every 2 weeks, for a total of three immunizations. (a) Detection of Anti-Pres1 antigen in serum; (b) Detection of Pres1 antigen in serum; (c) Detection of the level of HBV-associated antigen HBsAg in mouse serum. [Figure 11]Figure 11. Th cell helper epitopes enhanced antibody response to IFNα-Pres1-Fc vaccine. Compared with IFN-preS1-Fc, IFN-Pan-preS1-Fc can significantly enhance the immunogenicity of antigen molecules. C57 / BL6 (n=8 / group) mice were inoculated with aluminum-free hepatitis B Pres1, Pres1-Fc, and IFNα-Pres1-Fc proteins by subcutaneous immunization, and the levels of Pres1-specific antibodies in serum were detected by Elisa at the indicated times. [Figure 12] Figure 12. IFNα-Pan-Pres1-Fc was used as a therapeutic vaccine for chronic type B infection. C57 / BL6 mice were infected with 1x1011μg of AAV-HBV1.3 virus by tail vein injection, and 6 weeks after infection, stable infected mice (n=8 / group) were selected and inoculated with recombinant Pres1, IFNα-Pres1-Fc protein by subcutaneous immunization, once every 2 weeks, for a total of three immunizations. (a) Detection of Anti-Pres1 antigen in serum; (b) Detection of Pres1 antigen in serum; (c) Detection of the level of HBV-associated antigen HBsAg in mouse serum; (d) Detection of the level of HBV-DNA in mouse serum by QPCR. [Figure 13] Figure 13. IFNα-Pres1-Fc combined with HBsAg commercial vaccine can break the immune tolerance caused by HBsAg and induce HBsAg-HBsAb serological conversion. HBV Carrier mice were subcutaneously immunized with IFNα-Pres1-Fc and HBsAg commercial vaccine, once every 2 weeks, for a total of three times. (a) Levels of Pres1 in the serum of HBV Carrier mice; (b) levels of HBsAg; (c) levels of Anti-Pres1 in the serum; (d) levels of Anti-HBsAg in the serum; (e) levels of HBV-DNA in the serum. ***, p<0.001 [Figure 14]Figure 14. Compared with IFNα-RBD(SARS-CoV2)-Fc, free SARS-Cov2 RBD protein can elicit stronger antibody responses. Balb / c (n=8 / group) mice were inoculated with free SARS-Cov-2 RBD, RBD-Fc, and IFNα-RBD-Fc proteins by subcutaneous immunization, and the levels of SARS-Cov2 S protein-specific antibodies in serum were detected by Elisa at the indicated times. ****, p<0.0001. [Figure 15] Figure 15. After immunization of mice with IFNα-RBD(SARS-CoV2)-Fc, high-titer antiviral serum was produced, which could completely block SARS-CoV2 pseudovirus infection in in vitro cell experiments. [Figure 16] Figure 16. Detection of antiserum RBD-specific antibodies produced by immunization with IFNα-Pan-RBD (original strain)-Fc and IFNα-Pan-RBD (SARS-CoV-2 South African variant)-Fc. (a) Identification of IFNα-Pan-RBD (SARS-CoV-2 original strain)-Fc by SDS-PAGE electrophoresis. (b) Identification of IFNα-Pan-RBD (SARS-CoV-2 South African variant)-Fc by SDS-PAGE electrophoresis. (c) Combination of RBD-specific antibodies and original strain RBD in serum 14 days after immunization with IFNα-Pan-RBD (original strain)-Fc and IFNα-Pan-RBD (SARS-CoV-2 South African variant)-Fc. (d) RBD-specific antibodies in combination with South African variant RBD in serum 14 days after immunization with IFNα-Pan-RBD (original strain)-Fc and IFNα-Pan-RBD (SARS-CoV-2 South African variant)-Fc. [Figure 17] Figure 17. (a) Identification of Mouse IFNα-RBD-Fc and Mouse IFNα-Pan-RBD-Fc proteins by SDS-PAGE electrophoresis. (b) Identification of Human IFNα-RBD-Fc and Human IFNα-Pan-RBD-Fc proteins by SDS-PAGE electrophoresis. [Figure 18]Figure 18, Pan (Pan DR-binding epitope) CD4 T cell helper epitope can further enhance the immunogenicity of IFNα-RBD-Fc. Mice were inoculated with 10μg Mouse IFNα-RBD-Fc, Mouse IFNα-Pan-RBD-Fc or 10μg Human IFNα-RBD-Fc, Human IFNα-Pan-RBD-Fc protein by intramuscular immunization, and boosted 14 days after inoculation. Mouse serum was collected on days 7, 14, and 28 after immunization, respectively, and the levels of RBD-specific antibodies in mouse serum were detected by ELISA. *, p<0.05; ****, p<0.0001. [Figure 19] Figure 19. Aluminum adjuvant can enhance the specific humoral immune response induced by Human IFNα-RBD-Fc, Human IFNα-Pan-RBD-Fc proteins. C57BL / 6 mice were inoculated with 10 μg of Human IFNα-RBD-Fc or Human IFNα-Pan-RBD-Fc with or without aluminum adjuvant (AL+) on days 0 and 14, and mouse sera were collected on days 7, 14, and 28 after inoculation. The levels of SARS-CoV-2 RBD-specific antibodies in mouse sera were detected by ELISA. *, p<0.05; ****, p<0.0001. [Figure 20]Figure 20. IFN-Pan-RBD-Fc intranasal immunization can induce the production of high titers of RBD-specific IgG, IgA neutralizing antibodies. 6-8 week-old C57BL / 6 mice were divided into 5 groups with 10 mice per group and immunized with 10μg of IFNα-pan-RBD-Fc or equimolar amounts of RBD, RBD-Fc, and IFNα-RBD-Fc proteins by intranasal immunization, respectively, with the nasal drop volume being 10uL / mouse. Mice were immunized using two immunization procedures on days 0 and 14. Mouse serum was collected on days 7, 14, 21, 28, 35, and 42 after immunization, respectively, and the levels of novel coronavirus RBD-specific antibodies IgG (a) and IgA (b) in the serum of each group were detected by ELISA; serum on day 42 was collected and used for in vitro neutralization experiments of SARS-CoV-2 pseudovirus (c). Statistical method: one-way ANOVA, *p<0.05 significant difference, **p<0.01, ***p<0.001, ****p<0.0001. [Figure 21] Figure 21. Adjuvant-free SARS-CoV-2 vaccine IFN-Pan-RBD-Fc intranasal immunization induced the secretion of high titers of RBD-specific IgG and IgA neutralizing antibodies in nasopharyngeal and lung tissues. 28 days after immunization of 6-8 week-old C57BL / 6 mice, the mice were sacrificed and their nasal mucosa was collected and crushed with a tissue homogenizer. The homogenized liquid was centrifuged at 13,000 rpm for 10 minutes, and the supernatant was used as nasal mucosa supernatant (NMDS). ELISA was used to detect the levels of SARS-CoV-2 RBD-specific antibodies IgG (a) and IgA (b) in the nasal mucosa supernatant (NMDS) of each group; serum was collected on the 28th day and used for in vitro neutralization experiments of SARS-CoV-2 pseudovirus (c). Statistical method: one-way ANOVA, *p<0.05 significant difference, **p<0.01, ***p<0.001, ****p<0.0001. [Figure 22]Figure 22. Adjuvant-free novel coronavirus vaccine IFN-Pan-RBD-Fc intranasal immunization induced the secretion of high titers of RBD-specific IgG and IgA antibodies in lung tissue. 28 days after immunization (as shown in Figure 19), 6-8 week-old C57BL / 6 mice were sacrificed. In the lungs of mice, about 0.8 ml of HBSS+100 uM EDTA was aspirated with a 1 ml syringe, injected into the endotracheal tube, gently blown three times repeatedly, and then the liquid was aspirated and collected in a centrifuge tube. This operation was repeated three times, and finally about 2 ml of lung lavage fluid was obtained. The mouse lung lavage fluid was centrifuged at 500 g for 5 minutes, and the supernatant was used as mouse lung lavage fluid (BALF), and the novel coronavirus RBD-specific antibody IgG (a) and IgA (b) content in the mouse lung lavage fluid (BALF) of each group was detected by ELISA. Statistical method: one-way ANOVA, *p<0.05 significant difference, **p<0.01, ***p<0.001, ****p<0.0001. [Diagram 23] Figure 23. Expression and purification of Her2 vaccine proteins. The relevant proteins were expressed and purified in 293F cells, and the size and purity of the proteins were detected by SDS-PAGE and Coomassie brilliant blue staining. a. IFNα-3-Fc (62.6 kDa); b. IFNα-pan-3-Fc (63.9 kDa); c. IFNα-pan-4-Fc (74.9 kDa) and IFNα-4-Fc (73.6 kDa). [Figure 24] Figure 24. Analysis of antitumor activity of Her2 vaccine IFNα-3-Fc and IFNα-pan-3-Fc. TUBO breast cancer model mice were constructed, and the tumor size was 50-80 mm. Drug treatment was performed, and the relevant fusion proteins were injected intratumorally into the mice, once a week for a total of three treatments. The dosage was 10 μg / time / mouse for IFNα-3-Fc, equimolar for other fusion proteins, and CpG was used as an adjuvant. The tumor size was measured, and the tumor growth curve was plotted. [Diagram 25]Figure 25. Analysis of the function of IFNα and Pan to enhance the immunogenicity of Her2 antigen. BALB / C mice (n=5) aged 6-8 weeks were subcutaneously inoculated with mouse Her2 fusion protein vaccine 4-Fc, IFNα-4-Fc and IFNα-pan-4-Fc without the addition of adjuvant. The immunization dose was 10 μg / dose / mouse for IFNα-4-Fc, and equimolar for other fusion proteins. Venous blood was collected 14 and 21 days after immunization, and the level of specific IgG antibody in Her2 serum was detected by ELISA. [Figure 26] FIG. 26. Identification of IFN-HA1-Fc fusion protein by SDS-PAGE electrophoresis. [Figure 27] Figure 27. Mice were inoculated with 10ug of IFN by intramuscular immunization with tumor or equimolar amount of HA1 protein, and boosted 14 days after the first inoculation. Mouse serum was collected 28 days after immunization, and the level of HA1-specific antibody in mouse serum was detected by ELISA. On the 42nd day after immunization, mice were intranasally infected with 1000PFU of A / PR8 influenza virus, and the mice were observed from the 3rd day after virus infection, and the changes in body weight were recorded. (a) Mouse serum was collected 28 days after the first immunization, and the level of HA1-specific antibody in mouse serum was detected by ELISA. (b) Body weight changes of mice after virus infection. [Figure 28] Figure 28. Identification of IFNa-Pan-VZV-gE-Fc, IFNa-Pan-EBV-gp350-Fc, and IFNa-Pan-HSV-2-gD-Fc proteins by SDS-PAGE electrophoresis. DETAILED DESCRIPTION OF THE PREFERRED EMBODIMENTS

[0046] In order to make the object, technical solution and advantages of the present invention clearer, the present invention will be described in detail below with reference to the embodiments and the accompanying drawings. The described examples are only illustrative of the present invention and are not intended to limit the scope of the present invention, and the examples are only a part of the present invention and are not all examples of the present invention. The scope of the present invention is defined by the appended claims.

[0047] Example 1. Vaccine platform design The vaccine platform of interferon-target antigen-immunoglobulin Fc (or antibody) structural unit consisted of three structural units, the first structural unit was the interferon portion, the second structural unit was the immunoglobulin Fc region (or antibody), and the third structural unit was the target antigen. In the actual construction, the three structural units could be arranged and combined in any shape, and the target antigen could be linked to the Th cell helper epitope via linking sequence 2. The representative shape was as follows:

[0048] FIG. 1 was a schematic diagram of the vaccine platform, which combined the following order in the form of a homodimer: interferon-linked fragment 1-target antigen-immunoglobulin Fc.

[0049] FIG. 2 was a schematic diagram of the vaccine platform, in which the interferon-linked fragment 1-IgG1-hole and target antigen-IgG1-knob were combined in the form of heterodimers, respectively.

[0050] FIG. 3 was a schematic diagram of the vaccine platform, in which interferon-linked fragment 1-IgG1-knob and target antigen-IgG1-hole were combined in the form of a heterodimer.

[0051] The target antigen was then linked to a cellular helper epitope by linking fragment 2, which was then combined with the other two vaccine platform components, representative of which are as follows:

[0052] FIG. 4 is a schematic diagram of the vaccine platform, in which interferon-linked fragment 1-Th cell helper epitope-linked fragment 2-target antigen-immunoglobulin Fc are combined in the order in the form of a homodimer.

[0053] FIG. 5 was a schematic diagram of the vaccine platform combined in the form of heterodimers in the order of interferon-linked fragment 1-IgG1-hole and Th cell helper epitope-linked fragment 2-target antigen-IgG1-knob, respectively.

[0054] FIG. 6 was a schematic diagram of the vaccine platform, in which interferon-linked fragment 1-IgG1-knob and Th cell helper epitope-linked fragment 2-target antigen-IgG1-hole were combined in the form of heterodimers in this order.

[0055] Example 2. Construction, purification and production of vaccine platform We use the homodimer of Hepatitis B virus Pres1 and coronavirus SARS-CoV-2 RBD proteins as examples to illustrate the expression and production of this vaccine platform.

[0056] 1. Vector construction, host cell transfection and inducible expression 1.1. Using PEE12.4 as a vector, we constructed a vaccine structural unit into the vector by molecular cloning to obtain a plasmid capable of expressing the fusion protein, then transiently transfected 293F cells, collected the culture supernatant, and finally purified the target protein by protein A affinity chromatography column. Vector construction (containing HBV preS1 as an example) (1) PEE12.4-HindIII-signal peptide 1-interferon-BsiwI-Pres1-BstbI-hIgG1-EcoRI (2) PEE12.4-HindIII-signal peptide 1-interferon-BsiwI-RBD (SARS-CoV-2)-BstbI-hIgG1-EcoRI (3) PEE12.4-HindIII-signal peptide 1-interferon-Bsiwi-PADER-Pres1-hIgG1-EcoRI (4) PEE12.4-HindIII-signal peptide 1-interferon-Bsiwi-PADER-RBD (SARS-CoV-2)-hIgG1-EcoRI The linking sequences between each fusion protein fragment were as follows: (1) The linkage fragment 1 was between interferon and Pres1. (2) The linkage fragment 1 was between interferon and RBD (SARS-CoV-2). (3) The linking sequence between interferon and PADER was linking segment 1, and the linking segment between PADER and Pres1 was linking segment 2. (4) The linking sequence between interferon and PADER was linking segment 1, and the linking segment between PADER and RBD (SARS-CoV-2) was linking segment 2.

[0057] 1.2 Rapid expression of target proteins by transient transfection: (1) Cell resuscitation: 3 × 10 Freestyle 293F cells 7 The cells were frozen and stored in CD OptiCHOTM media (containing 10% DMSO) at a concentration of 1000 cells / ml. After removal from liquid nitrogen, the cells were quickly thawed in a 37°C water bath, added to a 15 ml centrifuge tube containing 10 ml of OptiCHOTM media, and centrifuged at 1000 rpm for 5 minutes. The supernatant was discarded, and the cell pellet was suspended in 30 ml of OptiCHOTM media and cultured at 37°C, 8% CO2, and 135 rpm. After 4 days, the cells were expanded, and the concentration of the expanded culture was 3 × 10 6 The cell density was not to exceed 100 cells / ml. (2) Two days before transfection, 0.6–0.8 × 10 6 Suspension culture 293F cells were prepared for transient transfection (200 ml) at a seeding density of 1000 cells / ml. (3) After 2 days, count the cell suspension to be transfected and determine the expected cell density of 2.5–3.5 × 10 6cells / ml, and the cell suspension was then centrifuged at 1000 rpm for 5 minutes and the supernatant discarded. (4) The cells were resuspended in 50 ml of fresh Freestyle 293 media and centrifuged again at 1,000 rpm for 5 minutes, and the supernatant was discarded. (5) 293F cells were resuspended in 200 ml of Freestyle 293 media. (6) 600 μg of plasmid was diluted with 5 ml of Freestyle 293 media, and bacteria were removed using a 0.22 μM filter. (7) 1.8 mg of PEI was diluted with 5 ml of Freestyle 293 media, and bacteria were removed using a 0.22 μM filter. Immediately after that, 5 ml of plasmid and 5 ml of PEI were mixed and left at room temperature for 5 minutes. (8) Add the plasmid / PEI mixture to the cell suspension, place in an incubator at 37°C, 8% CO2, and 85 rpm, and add 50 ug / L of growth factor LONG. TM The culture was carried out while supplementing with R3IGF-1. (9) After 4 hours, 200 ml of EX-CELLTM 293 media and 2 mM Glutamine were added, and the rotation speed was set to 135 rpm for continued cultivation. (10) After 24 hours, the cell growth inhibitor 3.8 mM VPA was added, and after 72 hours, 40 ml of medium D was added. After continued cultivation for 6 to 8 days (cell viability less than 70%), the supernatant was collected and left until it was used in the next purification step.

[0058] 1.3. Recovery, purification and electrophoretic validation of fusion protein

[0059] 2. Purification of target protein using Protein A: (1) Sample preparation: The suspension cell culture was transferred into a 500 ml centrifuge bucket and centrifuged at 8000 rpm for 20 min, the precipitate was discarded, and the supernatant was filtered through a 0.45 μM filter to remove impurities, after which NaN3 was added to a final concentration of 0.05% to prevent bacterial contamination during purification. Assembling the chromatography column: Take an appropriate amount of Protein A Agarose (calculated by purifying 20 mg of human Fc fusion protein per 1 ml of Protein A), mix well, add to the chromatography column, and leave at room temperature for about 10 minutes to overlay Protein A and 20% ethanol solution. Then, open the outlet at the bottom and allow the ethanol solution to slowly drain by gravity. (3) The chromatography column was washed with 10 column volumes of distilled water and binding buffer (20 mM sodium phosphate + 0.15 M NaCl, pH 7.0) and equilibrated. (4) Using a constant flow pump, the sample was loaded at a flow rate of 10 column volumes / hour, the flow-through was collected, and the sample loading was repeated twice. (5) The column was washed with at least 10 column volumes of binding buffer to remove impurities until no protein was detected in the effluent. (6) Elution was performed using Elution Buffer (0.1 M Glycine, pH 2.7), and each tube was collected in a separate tube (1 ml each), and the elution peak was observed using a protein indicator solution (Bio-Rad protein assay). The tubes containing the elution peaks were mixed and neutralized by adding an appropriate amount of 1 M Tris, pH 9.0 (pH was adjusted to 6-8, 0.5 or more higher than the isoelectric point of the purified protein). (7) Using a Zeba desalting and concentration centrifugal column, the target protein solution was replaced with the target buffer solution (the pH of the buffer solution was adjusted to avoid the isoelectric point of the protein). The protein concentration was determined by SDS-PAGE electrophoresis and NanoDrop2000 using BSA as a standard. (8) After elution, the column was washed successively with 20 column volumes of distilled water and 10 column volumes of 20% ethanol, and finally the ethanol solution was immersed in the gel medium and stored at 4°C.

[0060] 3. Identification of the above proteins by SDS-PAGE electrophoresis is shown in FIG.

[0061] Example 3. Compared with simple Pres1 antigen, FNα-Pres1-Fc, Pres1-Fc can induce stronger immune responses in mice.

[0062] Materials: C57BL / 6 male mice (5-8 weeks old) were purchased from Beijing Weitong Lihua Laboratory Animal Technology Co., Ltd., horseradish peroxidase (HRP)-labeled goat anti-mouse IgG was purchased from Beijing Kangwei Biological Technology Co., Ltd., 96-well ELISA measurement board was purchased from Corning Costar, ELISA color development solution was purchased from eBioscience, and the microplate reader SPECTRA max PLUS 384 used was purchased from Molecular, USA. The aluminum adjuvant used was purchased from SIGMA.

[0063] method: (1) Immunization of mice with Pres1 fusion protein: 80 pmol of IFN-Pres1-Fc or 80 pmol of Pres1-Fc, Pres1 protein was mixed with aluminum adjuvant and then subcutaneously immunized. Antibody detection was performed by collecting mouse sera by orbital bleeding at the indicated time points. (2) Antibodies produced by IFNα-Pres1-Fc have a broad neutralizing effect against HBV viruses of different genotypes. Five-week-old male C57BL / 6 mice were inoculated with AAV-HBV 1.3 (HBV genotypes B, C, and D) at 1x10 11 1 μg virus was infected, and 6 weeks later, mice with stable expression of HBV antigen were screened and tested. The screened mice (4 mice / group) were intravenously injected with 200 ul / mouse of serum from IFNα-Pres1-Fc-immunized mice. After 12 hours, mouse serum was collected, and changes in mouse Pres1 antigen before and after antiserum administration were detected by Elisa. (3) Detection of anti-Pres1 specific antibodies in serum by ELISA. Pres1 (2ug / ml) coating solution was added to Elisa plates (Corning 9018) at 50ul / well and coated overnight at 4℃. The plates were washed once with 260ul PBS per well. They were blocked with 5% blocking solution (5% FBS) at 37℃ for 2 hours. Serum samples (1:10, 1:100, 1:1000, 1:10000) were diluted in PBS and added to the blocked Elisa plates at 50ul / well and incubated at 37℃ for 1 hour. After washing five times with 260ul PBST each time, 50ul of enzyme-conjugated secondary antibody (enzyme-conjugated anti-mouse IgG-HRP 1:5000 diluted by PBS) was added to each well and incubated at 37℃ for 1 hour. Wash five times with 260ul PBST each time, add substrate TMB at 100ul / well, incubate at room temperature away from light, wait for the substrate to develop, add 50ul of termination solution (2N H2SO4) to each well to terminate the color development, read the plate with a microplate reader, OD450-630.

[0064] Results: Free Pres1 had weak immunogenicity, but the addition of IFNα and Fc moieties to Pres1 to form IFNα-Pres1-Fc fusion protein significantly improved its immunogenicity, as shown in Figure 8(a). Moreover, the antibodies produced by IFNα-Pres1-Fc had a broad neutralizing effect against different HBV genotypes, as shown in Figure 8(b).

[0065] Example 4. IFNα-Pres1-Fc can be used as a prophylactic vaccine against Hepatitis B. Materials: C57BL / 6 (6-8) week old male mice were purchased from Beijing Weitong Lihua Biotechnology Co., Ltd., HBsAg detection kit was purchased from Shanghai Kehua Biotechnology Co., Ltd. AAV-HBV 1.3 virus was purchased from Guangzhou Paijin Biotechnology Co., Ltd. Other experimental materials were the same as in Example 3.

[0066] method: (1) Mice were subcutaneously immunized with 80 pmol of different forms of Pres1 vaccines, including simple Pres1, Pes1-Fc, and IFNα-Pres1-Fc proteins. Serum was collected 28 days after immunization and then diluted with 1x10 11 Mice were infected with 1 μg AAV-HBV 1.3 virus and serum samples were taken every week for 4 consecutive weeks to detect anti-Pres1 antibodies, HBsAg, and Pres1 antigen in the serum. At the third week, the levels of HBV-DNA in the periphery of the mice were detected. (2) Detection of Pres1 specific antigen in serum by ELISA Antigen coating: Pres1 antibody XY007 (4ug / ml) coating solution was added to Elisa plate (Corning 9018) at 50ul / well and coated at 4℃ overnight. Plate was washed once with 260ul PBS per well. Blocked with 5% blocking solution (5%FBS) at 37℃ for 2 hours. Serum samples (1:10, 1:100) were diluted with PBS and added to the blocked Elisa plate at 50ul / well, setting up two replicate wells per dilution, and incubated at 37℃ for 1 hour. Washed five times with 260ul PBST each time, and added 50ul enzyme conjugate (from KH HBsAg Detection Kit) to each well and incubated at 37℃ for 1 hour. Wash five times with 260ul PBST each time, add substrate TMB 100ul / well, incubate at room temperature away from light, wait for the substrate to develop, add 50ul of termination solution (2N H2SO4) to each well to terminate the color development, read the plate with enzyme marker, OD450-630.

[0067] Results: As shown in Figure 9(a), mice in the IFNα-Pres1-Fc immunized group were able to produce high levels of Pres1 antibodies before virus inoculation and continued to maintain high levels during virus infection. Compared with the group without protein immunization, IFN-Pres1-Fc vaccine immunization could significantly prevent HBV infection, and the anti-preS1 antibodies produced after immunization could rapidly and completely eliminate preS1 antigen in serum (Figure 9(b)), and most of the mice infected with virus in the IFN-Pres1-Fc immunized group showed peripheral HBsAg negativity (Figure 9(c, d)). The above experimental results, as shown in Figure 9, indicate that IFN-Pres1-Fc as a vaccine can effectively prevent HBV infection.

[0068] Example 5. IFNα-Pres1-Fc was used as a therapeutic vaccine for chronic type B infection. Materials: C57BL / 6 male mice (4 weeks old) were purchased from Beijing Weitong Lihua Biological Technology Co., Ltd. AAV-HBV 1.3 was purchased from Guangzhou Paijin Biological Technology Co., Ltd. HBsAg detection kit was purchased from Shanghai Kehua Biological Technology Co., Ltd. Other experimental materials were the same as in Example 4.

[0069] method: (1) Screening of HBV carrier mice: 4-week-old C57BL / 6 mice were injected with 1x10 11 The mice were injected with 1 μg AAV-HBV 1.3 virus, and the HBV antigen HBsAg was detected 1 to 6 weeks later. Mice with stable expression of HBsAg were screened and used as HBV carrier mice. (2) The screened mice were subcutaneously injected with 80 pmol of different forms of Pres1 protein once every 2 weeks for a total of three immunizations. Mouse serum was collected 14 days after immunization and then once a week, and the levels of anti-Pres1 antibody, HBsAG, and Pres1 antigen in mouse serum were detected by ELISA. After the final blood collection, the HBV-DNA content in the peripheral blood of the mice was detected.

[0070] Results: The changes in serum preS1 antigen, serum Pres1 antibody and HBsAg of carrier mice immunized with IFN-Pres1-Fc vaccine were detected. The results show that after IFNα-Pres1-Fc vaccine immunization, mice produced high levels of anti-Pres1 antibody (as shown in Figure 10(a)), and preS1 antigen in serum could be completely eliminated (as shown in Figure 10(b)). At the same time, serum HBsAg was also reduced to a certain extent (as shown in Figure 10(c)), but there was no therapeutic effect in both the untreated control group and the simple Pres1 vaccine immunization group (as shown in Figure 10).

[0071] Example 6. T cell helper epitopes enhanced antibody responses to IFNα-Pres1-Fc vaccine. The materials were the same as in Example 3.

[0072] method: (1) Immunization of mice with Pres1 fusion proteins: Mice were subcutaneously immunized with 80 pmol of IFN-Pan-Pres1-Fc or 80 pmol of IFN-Pan-Pres1-Fc, Pres1-Fc, or Pres1 proteins containing the Pan epitope. Antibody detection was performed by collecting mouse sera by retroorbital bleeding at the indicated time points. (2) Detection of anti-Pres1 specific antibodies in serum by ELISA, same as in Example 3.

[0073] Results:Compared with fusion protein vaccines such as IFN-preS1-Fc, IFN-Pan-preS1-Fc can significantly enhance the immunogenicity of antigen molecules and induce the production of broadly neutralizing antibodies.C57 / BL6 (n=8 / group) mice were inoculated with aluminum-free hepatitis B Pres1, Pres1-Fc, and IFNα-Pres1-Fc proteins by subcutaneous immunization, and the levels of Pres1-specific antibodies in serum were detected by Elisa at the designated times.

[0074] Example 7 IFNα-Pan-Pres1-Fc was used as a prophylactic vaccine against chronic type B infection. Materials: C57BL / 6 male mice (4 weeks old) were purchased from Beijing Weitong Lihua Biological Technology Co., Ltd. AAV-HBV 1.3 was purchased from Guangzhou Paijin Biological Technology Co., Ltd. HBsAg detection kit was purchased from Shanghai Kehua Biological Technology Co., Ltd. Other experimental materials were the same as in Example 4.

[0075] method: (1) Screening of HBV carrier mice: 4-week-old C57BL / 6 mice were injected with 1x10 11 The mice were injected with 1 μg AAV-HBV 1.3 virus, and the HBV antigen HBsAg was detected 1 to 6 weeks later. Mice with stable expression of HBsAg were screened and used as HBV carrier mice. (2) The screened mice were subcutaneously injected with 80 pmol of different forms of Pres1 protein once every 2 weeks for a total of three immunizations. Mouse serum was collected 14 days after immunization and then once a week, and the levels of anti-Pres1 antibody, HBsAG, and Pres1 antigen in mouse serum were detected by ELISA. After the final blood collection, the HBV-DNA content in the peripheral blood of the mice was detected.

[0076] Results: We detected the changes in serum preS1 antigen, serum Pres1 antibody and HBsAg in carrier mice immunized with IFN-Pan-Pres1-Fc vaccine. The results show that after IFN-Pan-Pres1-Fc vaccine immunization, the mice produced high levels of anti-Pres1 antibody (as shown in Figure 12(a)). And the preS1 antigen in serum could be completely removed (as shown in Figure 12(b)), and HBsAg in serum was also reduced to a certain extent (as shown in Figure 12(c)), but there was no therapeutic effect in either the untreated control group or the simple Pres1 vaccine immunization group. And the HBV DNA was also significantly reduced in the IFNα-Pan-Pres1-Fc immunization group (as shown in Figure 12(d)).

[0077] Example 8. IFNα-Pan-Pres1-Fc can be combined with HBsAg commercial vaccine to break the immune tolerance caused by HBsAg and induce HBsAg-HBsAb serological conversion. Materials: C57BL / 6 male mice (4 weeks old) were purchased from Beijing Weitong Lihua Biological Technology Co., Ltd. AAV-HBV 1.3 was purchased from Guangzhou Paijin Biological Technology Co., Ltd. HBsAg detection kit was purchased from Shanghai Kehua Biological Technology Co., Ltd. Anti-HBsAg kit was purchased from Beijing Wantai Biological Pharmaceutical Co., Ltd. Commercially available HBsAg vaccine was purchased from Emei Hanxin Vaccine (Dalian) Co., Ltd. Other experimental materials were the same as in Example 7.

[0078] method: (1) Screening of HBV carrier mice: 4-week-old C57BL / 6 mice were injected with 1x10 11 The mice were injected with 1 μg AAV-HBV 1.3 virus, and the HBV antigen HBsAg was detected 1 to 6 weeks later. Mice with stable expression of HBsAg were screened and used as HBV carrier mice. (2) Screened HBV carrier mice were immunized with 80 pmol of IFNα-pan-Pres1-Fc and 2 μg of a commercially available HBsAg vaccine twice consecutively at an interval of 14 days. Mouse serum was collected 14 days after the first immunization, and then every week to detect changes in anti-Pres1, Pres1, anti-HBsAg, and HBsAg in the mouse serum. The level of HBV-DNA in the serum was then detected when the mouse serum was finally collected.

[0079] Results: Combining IFNα-Pan-Pres1-Fc with commercial HBsAg as a therapeutic strategy for chronic hepatitis B can eventually break the immune tolerance caused by HBsAg. The immune response generated in HBV-resistant mice can completely eliminate preS1 antigen in serum (as shown in Figure 13(a)), and there is a high concentration of Pres1 antibody in serum (as shown in Figure 13(c)). What is exciting is that IFN-Pan-Pres1-Fc vaccine can effectively eliminate HBsAg in serum while inducing partial conversion of serological HBsAb (as shown in Figures 13(b) and 14(d)). This is considered to be a clinically significant indicator of HBV cure. In addition, the expression levels of HBV-related DNA in peripheral blood were detected by fluorescent quantitative PCR (real-time PCR), and the results showed that the immunization method combining IFNα-Pan-Pres1-Fc and commercially available HBsAg could ultimately reduce peripheral HBV DNA levels compared to the control group (as shown in Figure 13(e)). Based on these results, we invented a vaccine strategy for the treatment of chronic hepatitis B using a vaccine combining IFNα-Pan-Pres1-Fc and commercially available HBsAg.

[0080] Example 9. Compared to IFNα-RBD(SARS-CoV2)-Fc, free SARS-Cov2 RBD protein can elicit stronger antibody responses. Materials: Balb / c male and female mice (6-8 weeks) were purchased from Beijing Weitong Lihua Laboratory Animal Technology Co., Ltd., and the SARS-CoV-2 RBD protein used was purchased from Beijing Keyue Zhongkai Biological Technology Co., Ltd. 293-hACE2 cells were provided by Professor Zhang Zheng (Shenzhen Third People's Hospital). Luciferase Reporter detection kit was purchased from Promega. Other experimental materials were the same as in Example 3.

[0081] method: (1) Immunization of mice with IFNα-RBD (SARS-Cov-2)-Fc fusion protein: 10ug of IFNα-RBD-Fc, RBD-Fc or 10ug of RBD protein was mixed with aluminum adjuvant and then immunized subcutaneously. 28 days after immunization, mouse serum was collected by orbital bleeding to detect novel coronavirus-specific antibodies. (2) Detection of SARS-cov2 RBD antibodies in serum. Antigen coating: RBD (1.5ug / ml) coating solution was added to Elisa plates (Corning 9018) at 100ul / well and coated overnight at 4℃. Plates were washed once with 260ul PBS per well. Blocked with 100ul 5% blocking solution (5%FBS) at 37℃ for 2 hours. Serum samples (1:10, 1:100, 1:1000, 1:10000, 1:100000…) were diluted in PBS and added to the blocked Elisa plates at 100ul / well and incubated at 37℃ for 1 hour. Washed 5 times with 260ul PBST each time, 100ul of enzyme-conjugated secondary antibody (enzyme-conjugated anti-mouse IgG-HRP 1:5000 diluted by PBS) was added to each well and incubated at 37℃ for 1 hour. After washing twice with 260ul of PBST, 100ul / well of substrate TMB was added and incubated at room temperature for 15 minutes away from light to allow the substrate to develop color. 50ul of termination solution (2N H2SO4) was added to each well to terminate color development, and the plate was read with a microplate reader, with OD450-630. Titer calculation method: The maximum dilution factor that gave a positive result was selected, and the OD value / Cutoff value (0.1) corresponding to that dilution factor was multiplied by the dilution factor (X) to obtain the antibody titer corresponding to that serum. (3) In vitro neutralization experiment of SARS-CoV-2 protein pseudovirus Antiserum was diluted 1:3 and added to a 96-well plate, 50ul of pseudovirus particles containing luciferase spike protein was added to the well, the virus-antibody mixture was left at 37℃ for 1 hour, and 293-hACE2 cells were added to the 96-well plate at 10^4 / well. The 96-well plate was placed in a cell culture incubator at 37℃, and luciferase activity was detected after 48 hours.

[0082] Results: The immunogenicity of the free novel coronavirus was weak, and when IFNα and Fc moieties were added to the novel coronavirus RBD polypeptide protein region to form IFNα-RBD-Fc fusion protein, its immunogenicity was significantly improved, as shown in Figure 14. And the antibodies produced by IFNα-RBD-Fc could block the infection of cells by the pseudovirus of SARS-CoV-2 S protein in vitro, as shown in Figure 15.

[0083] Example 10. Detection of antiserum RBD-specific antibodies produced by immunization with IFNα-Pan-RBD (original strain)-Fc and IFNα-RBD (SARS-CoV-2 South African variant)-Fc. Materials: Balb / c male and female mice (6-8 weeks) were purchased from Beijing Weitong Lihua Laboratory Animal Technology Co., Ltd., and the SARS-CoV-2 original strain RBD protein used was purchased from Beijing Keyue Zhongkai Biological Technology Co., Ltd. The SARS-CoV-2 South African mutant strain RBD protein used was purchased from Sino Biological, Inc. Other experimental materials were the same as in Example 3.

[0084] method: (1) The construction and expression methods of IFNα-Pan-RBD (original strain)-Fc and IFNα-RBD (SARS-CoV-2 South African variant)-Fc proteins were the same as in Example 2. (2) Immunization of mice with IFNα-Pan-RBD (original strain)-Fc and IFNα-Pan-RBD (SARS-CoV-2 South African variant)-Fc fusion proteins: 10ug of IFNα-Pan-RBD (original strain)-Fc or IFNα-Pan-RBD (SARS-CoV-2 South African variant)-Fc protein was mixed with aluminum adjuvant and then immunized subcutaneously. 14 days after immunization, mouse serum was collected by orbital blood sampling to detect novel coronavirus-specific antibodies. (3) Analysis of antibody responses by ELISA was the same as in Example 9.

[0085] Results: The SDS-PAGE results showed the accurate band size of IFNα-Pan-RBD (SARS-CoV-2 original strain)-Fc, indicating that the mutant novel coronavirus IFNα-RBD (SARS-CoV-2 original strain)-Fc vaccine protein was successfully constructed, expressed and purified (Figure 16a). The SDS-PAGE results showed the accurate band size of IFNα-Pan-RBD (SARS-CoV-2 South African variant)-Fc, indicating that the mutant novel coronavirus IFNα-RBD (SARS-CoV-2 South African variant)-Fc vaccine protein was successfully constructed, expressed and purified (Figure 16b). The ELISA results show that the antibodies produced by induction in IFNα-Panan-RBD (original strain)-Fc immunized mice, as well as in IFNα-Pan-RBD (SARS-CoV-2 South African variant)-Fc immunized mice, were all able to bind to the original strain of the new coronavirus RBD protein, and there was no significant difference in the binding ability of the induced antibodies from both strains to the original strain RBD (Figure 16c). The ELISA results for the South African variant RBD show that the antibodies produced by induction in IFNα-Panan-RBD (original strain)-Fc immunized mice, as well as in IFNα-Pan-RBD (SARS-CoV-2 South African variant)-Fc immunized mice, were all able to bind to the South African variant RBD, and the binding ability was equivalent (Figure 16c).

[0086] Example 11. Materials: C57BL / 6 female mice (6-8 weeks) were purchased from Beijing Weitong Lihua Biological Technology Co., Ltd. The SARS-CoV-2 RBD protein used in the ELISA was purchased from Beijing Keyue Zhongkai Biological Technology Co., Ltd. The Mouse IFNα-RBD-Fc, Mouse IFNα-Pan-RBD-Fc, Human IFNα-RBD-Fc, and Human IFNα-Pan-RBD-Fc proteins used for immunization were all produced in our laboratory, and other experimental materials were the same as those in Example 3.

[0087] method: (1) The design of the fusion protein, the construction of the plasmid, and the method of purifying the protein were the same as in Examples 1 and 2. (2) Immunization of mice with vaccine proteins 10μg of Mouse IFNα-RBD-Fc, Mouse IFNα-Pan-RBD-Fc or 10μg of Human IFNα-RBD-Fc, Human IFNα-Pan-RBD-Fc vaccine proteins were mixed with 20μg of aluminum adjuvant overnight, and then vaccinated into mice by intramuscular immunization. A booster immunization was performed 14 days after the first vaccination. Mouse sera were collected on days 7, 14, and 28 after immunization, and the levels of RBD-specific antibodies in the mouse sera were detected by ELISA. (3) Detection of SARS-cov2 RBD antibodies in serum Antigen coating: RBD (1.5ug / ml) coating solution was added to Elisa plates (Corning 9018) at 100ul / well and coated overnight at 4℃. Plates were washed once with 260ul PBS per well. Blocked with 100ul 5% blocking solution (5%FBS) at 37℃ for 2 hours. Serum samples (1:10, 1:100, 1:1000, 1:10000, 1:100000…) were diluted in PBS and added to the blocked Elisa plates at 100ul / well and incubated at 37℃ for 1 hour. Washed 5 times with 260ul PBST each time, 100ul of enzyme-conjugated secondary antibody (enzyme-conjugated anti-mouse IgG-HRP 1:5000 diluted by PBS) was added to each well and incubated at 37℃ for 1 hour. After washing with 260ul of PBST, 100ul of substrate TMB was added to each well, and incubated at room temperature for 15 minutes away from light to allow the substrate to develop color. 50ul of termination solution (2N H2SO4) was added to each well to terminate the color development, and the plate was read with an enzyme marker, with OD450-630. Titer calculation method: The maximum dilution factor that gave a positive result was selected, and the OD value / Cutoff value (0.1) corresponding to that dilution factor multiplied by the dilution factor (X) was used as the antibody titer corresponding to the serum.

[0088] result: As shown in Figure 17, after the protein was expressed and purified, the SDS-PAGE results showed that the protein size was as expected and a single band appeared at the target position. As shown in Figure 18, the addition of the Pan (Pan DR-binding epitope) CD4 T cell helper epitope can enhance the immunity of Mouse IFNα-RBD-Fc and Human IFNα-RBD-Fc. The experimental results show that the addition of the pan epitope results in higher RBD-specific antigen production in both Mouse IFNα-Pan-RBD-Fc and Human IFNα-Pan-RBD-Fc compared to Mouse IFNα-RBD-Fc, regardless of whether it is 7 days, 14 days, or 28 days after immunization with the vaccine protein.

[0089] Example 12. Materials: C57BL / 6 female mice (6-8 weeks old) were purchased from Beijing Weitong Lihua Biological Technology Co., Ltd., and the SARS-CoV-2 RBD protein used in the ELISA was purchased from Beijing Keyu Zhongkai Biological Technology Co., Ltd. The Human IFNα-RBD-Fc and Human IFNα-Pan-RBD-Fc proteins used for immunization were both produced in our laboratory. Other experimental materials were the same as in Example 3.

[0090] method: (1) Immunization of mice with Human IFNα-RBD-Fc and Human IFNα-Pan-RBD-Fc proteins: 10ug of Human IFNα-RBD-Fc or Human IFNα-Pan-RBD-Fc proteins were mixed with aluminum adjuvant overnight to prepare vaccine samples containing aluminum adjuvant, whereas 10ug of Human IFNα-RBD-Fc or Human IFNα-Pan-RBD-Fc proteins were diluted with PBS to prepare vaccine samples without aluminum adjuvant. Mice were inoculated intramuscularly with 10μg of Human IFNα-RBD-Fc or Human IFNα-Pan-RBD-Fc proteins in the presence or absence of aluminum adjuvant, and boosted 14 days after inoculation. Mouse sera were collected on days 7, 14, and 28 after immunization, and the levels of RBD-specific antibodies in the mouse sera were detected by ELISA. (2) Detection of SARS-cov2 RBD antibodies in serum. Antigen coating: RBD (1.5ug / ml) coating solution was added to Elisa plates (Corning 9018) at 100ul / well and coated overnight at 4℃. Plates were washed once with 260ul PBS per well. Blocked with 100ul 5% blocking solution (5%FBS) at 37℃ for 2 hours. Serum samples (1:10, 1:100, 1:1000, 1:10000, 1:100000…) were diluted in PBS and added to the blocked Elisa plates at 100ul / well and incubated at 37℃ for 1 hour. Washed 5 times with 260ul PBST each time, 100ul of enzyme-conjugated secondary antibody (enzyme-conjugated anti-mouse IgG-HRP 1:5000 diluted by PBS) was added to each well and incubated at 37℃ for 1 hour. After washing with 260ul of PBST, 100ul of substrate TMB was added to each well, and incubated at room temperature for 15 minutes away from light to allow the substrate to develop color. 50ul of termination solution (2N H2SO4) was added to each well to terminate the color development, and the plate was read with an enzyme marker, with OD450-630. Titer calculation method: The maximum dilution factor that gave a positive result was selected, and the OD value / Cutoff value (0.1) corresponding to that dilution factor multiplied by the dilution factor (X) was used as the antibody titer corresponding to the serum.

[0091] result: As shown in Figure 19, the application of aluminum adjuvant can enhance the immunogenicity of Human IFNα-RBD-Fc and Human IFNα-Pan-RBD-Fc proteins. Without adjuvant, Human IFNα-RBD-Fc and Human IFNα-Pan-RBD-Fc vaccines can generate high titer antibody responses, but Human IFNα-RBD-Fc and Human IFNα-Pan-RBD-Fc proteins adjuvanted with aluminum adjuvant can further increase the level of RBD-specific antibody responses on days 7, 14, and 28 after vaccination compared to the adjuvanted group without adjuvant.

[0092] Example 13. material: The experimental animals were purchased from Beijing Weitong Lihua Laboratory Animal Co., Ltd., and the animals used were 6-8 week-old C57BL / 6 mice; Animal certificate number: No. 110011200106828974; RBD protein for immunization was purchased from Beijing Keyu Zhongkai Biological Technology Co., Ltd., and RBD-Fc, IFNα-RBD-Fc and IFN-pan-RBD-Fc proteins were produced in our laboratory; All adjuvants were purchased from SERVA, Germany; Horseradish peroxidase (HRP)-labeled goat anti-mouse IgG was purchased from Beijing Kangwei Biological Technology Co., Ltd.; 96-well ELISA measurement board was purchased from Corning Costar; ELISA color development solution was purchased from eBioscience; The microplate reader SPECTRA max PLUS 384 used was purchased from Molecular, USA; Tissue homogenizer was purchased from Beijing Haonuos Technology Co., Ltd.

[0093] method: Mice aged 6 to 8 weeks were divided into 5 groups with 10 mice per group and immunized with 10ug of IFNα-pan-RBD-Fc or equimolar amounts of RBD, RBD-Fc, and IFNα-RBD-Fc proteins by intranasal immunization, respectively, with the nasal drop volume being 10uL / mouse. Mice were immunized using two immunization procedures on days 0 and 14. Mouse serum was collected on days 7, 14, 21, 28, 35, and 42 after immunization, respectively, and the content of SARS-CoV-2 RBD-specific antibodies in the serum of each group was detected by ELISA; serum on day 28 was collected and used for in vitro neutralization experiments of SARS-CoV-2 pseudovirus.

[0094] result: As shown in Figure 20, two intranasal immunizations of RBD, RBD-Fc protein can induce a certain degree of antibody response, and the levels of both serum IgG and IgA induced at the same time point after two intranasal immunizations of IFNα-pan-RBD-Fc were significantly higher than those in the RBD, RBD-Fc and IFN-RBD-Fc groups, and the results of the pseudovirus neutralization experiment indicate that IFN-RBD-Fc can induce a higher level of neutralizing antibody production compared with the RBD and RBD-Fc immunization groups.

[0095] Example 14. material: Same as Example 10.

[0096] method: Mice aged 6 to 8 weeks were divided into 4 groups, 5 mice per group, and immunized with 10ug of IFNα-pan-RBD-Fc or equimolar amounts of RBD, RBD-Fc, and IFNα-RBD-Fc proteins, respectively, by intranasal immunization, with a dose of 10uL / mouse. Mice were immunized using two immunization procedures on days 0 and 14. On the 28th day after immunization, the nasal mucosa supernatants and lung lavage fluids of the mice were collected, and the content of SARS-CoV-2 RBD-specific antibodies in the serum of each group was detected by ELISA, and the SARS-CoV-2 pseudovirus neutralization experiment was used to detect the novel coronavirus pseudovirus in the serum and nasal mucosa supernatant. Obtaining mouse nasal mucosa supernatant and lung lavage fluid from immune experimental animals: Mice were put to sleep and killed, and the mouse nasal mucosa was collected and crushed with a tissue homogenizer. The homogenized liquid was centrifuged at high speed at 13000 rpm for 10 minutes, and the supernatant was used as nasal mucosa supernatant (NMDS). For mouse lungs, about 0.8 ml of HBSS + 100 uM EDTA was aspirated with a 1 ml syringe and injected into an endotracheal tube, gently blown three times, and then the liquid was aspirated and collected in a centrifuge tube. This operation was repeated three times, and finally about 2 ml of lung lavage fluid was obtained. The mouse lung lavage fluid was centrifuged at 500 g for 5 minutes, the supernatant was used as mouse lung lavage fluid (BALF), and the precipitate was lymphocytes in the mouse lung, which could be further analyzed.

[0097] result: As shown in Figure 21, IFNα-pan-RBD-Fc protein can induce strong local IgG antibody response and IgA mucosal immunity in the nasal mucosa by two intranasal immunizations compared with RBD and RBD-Fc protein. The strength of IFNα-pan-RBD-Fc protein response was stronger than that of RBD and RBD-Fc groups. The results of pseudovirus neutralization experiments show that IFNα-pan-RBD-Fc protein immunization group can induce the production of higher titer neutralizing antibodies in the nasal mucosa. As shown in Figure 22, two intranasal immunizations of C57BL / 6 mice with IFNα-pan-RBD-Fc fusion protein induced similar potent IgG and IgA antibody secretion in local lung tissues. The results of pseudovirus neutralization experiments showed that IFNα-pan-RBD-Fc induced higher titers of neutralizing antibodies than RBD, RBD-Fc.

[0098] Example 15. Her2 belongs to the HER family of type I transmembrane growth factor receptors and consists of an extracellular ligand-binding domain, a transmembrane domain and an intracellular tyrosine kinase domain. When a ligand binds to the extracellular domain, the HER protein dimerizes and transphosphorylates its intracellular domain, and the phosphorylated tyrosine residues bind to various intracellular signaling molecules, activate downstream signaling pathways and regulate gene transcription. Most of the regulated genes are related to cell proliferation, survival, differentiation, angiogenesis, invasion and metastasis. The extracellular segment of the Her2 protein is relatively large, containing more than 600 amino acids, and can be divided into four structural domains, namely domains I, II, III and IV. The currently approved Trastuzumab mainly binds to domain IV, Pertuzumab mainly binds to domain II, and the peptide vaccine E75 in clinical trials targets domain III. There are several important sites in different structural domains, indicating that they may mediate antitumor effects. To research vaccine platforms for tumor prevention and treatment, this patent investigated and selected the tumor antigen Her2 as a target, constructed IFN-Her2-Fc and IFN-Pan-Her2-Fc to construct fusion protein vaccines, and analyzed the in vivo antitumor activity and vaccine immune activity.

[0099] material and method: material: BALB / c female mice (6-8 weeks) were purchased from Beijing Weitong Lihua Biotechnology Co., Ltd., TUBO cells were from TCGA, and other materials were the same as in Example 3. method: (1) The design of the fusion protein, the construction of the plasmid, and the method of purifying the protein were the same as in Examples 1 and 2. First, we constructed expression plasmids for domains III and IV of the extracellular segment of mouse Her2 (designated IFNα-3-Fc, IFNα-pan-3-Fc, IFNα-pan-4-Fc and IFNα-4-Fc, respectively), and expressed and purified the relevant proteins in human 293F cell line. The size and purity of the proteins were identified by SDS-PAGE and Coomassie Brilliant Blue staining. (2) Analysis of the direct antitumor activity of IFNα-3-Fc and IFNα-pan-3-Fc TUBO is a breast cancer cell line derived from BALB-NeuT mice and has been used to study the growth and treatment of Her2-positive breast cancer. TUBO tumors were used to detect the antitumor activity of IFNα in the protein. TUBO breast cancer model mice were constructed, and 5*105 TUBO cells were subcutaneously inoculated into BALB / C mice, with tumor sizes ranging from 50 to 80 mm3. The mice were treated once a week for a total of three doses. The doses of IFNα-3-Fc were 10 μg / mouse, the other drugs were administered equimolarly, and CpG was used as an adjuvant. The tumor size was measured, and the tumor growth curve was plotted. (3) Analysis of the improvement of immunogenicity of Her2 vaccine by IFNα and Pan BALB / C female mice aged 6 to 8 weeks were subcutaneously inoculated once a week with the adjuvant-free HER2 domain V fusion protein vaccine 4-Fc, IFNα-4-Fc, and IFNα-pan-4-Fc for a total of three times. The immunization dose was 10 μg / mouse for IFNα-4-Fc, and equimolar doses for the other proteins. Venous blood was collected 14 and 21 days after immunization, and the levels of Her2-specific IgG antibodies were detected by ELISA.

[0100] result: (1) As shown in Figure 23, the size of the Her2 fusion proteins was almost as expected, and the purity also met the experimental requirements: IFNα-3-Fc (62.6 kDa), IFNα-pan-3-Fc (63.9 kDa), IFNα-pan-4-Fc (74.9 kDa) and IFNα-4-Fc (73.6 kDa). Under non-denaturing conditions, the proteins were in a dimeric state, consistent with autodimerization of the Fc fragment. (2) As shown in Figure 24, compared with the control group, intratumoral injection of Her2 fusion proteins IFNα-pan-3-Fc and IFNα-3-Fc significantly inhibited the growth of TUBO tumors, and the control effect was similar to that of the IFNα-Fc group. This indicates that IFNα activity is good in protein vaccines and there are no factors such as steric hindrance that affect IFNα activity, which can be used to further explore its efficacy and mechanism in anti-tumor immunity. (3) As shown in Figure 25, compared with the control group, 4-Fc, IFNα-4-Fc, and IFNα-pan-4-Fc could induce obvious Her2-specific IgG antibody responses 14 and 21 days after Her2 fusion protein vaccine immunization, and compared with 4-Fc, the antibody titers induced by IFNα-4-Fc and IFNα-pan-4-Fc tended to be higher. Also, on the 21st day after immunization, the antibody titers induced by IFNα-pan-4-Fc were significantly higher than those of the 4-Fc group. This indicates that the addition of IFNα and pan contributed to the increase in the immunogenicity of 4-Fc and induced a stronger antigen-specific antibody response, and therefore IFN-Pan-HER2-Fc and IFN-Pan-HER2-Fc were potentially effective tumor vaccines against Her2-positive tumors.

[0101] Example 16. Materials: BALB / c female mice (6-8 weeks) were purchased from Beijing Weitong Lihua Biological Technology Co., Ltd.; HA1 (A / PR8) protein used in ELISA was purchased from Sino Biological, Inc.; HA1 protein (A / PR 8) for immunization was purchased from Sino Biological, Inc.; IFNα-HA1-Fc was produced in our laboratory; H1N1 (A / PR8) influenza virus used to infect mice was produced in our laboratory; other experimental materials were the same as in Example 3.

[0102] method: (1) The design of IFNα-HA1-Fc protein, the construction of the plasmid, and the purification of the protein were the same as in Examples 1 and 2. (2) Immunization of mice with HA1 and IFNα-HA1-Fc proteins. 10 μg of IFNα-HA1-Fc or an equimolar amount of HA1 protein was mixed with 20 μg of aluminum adjuvant overnight, and then inoculated into mice by intramuscular immunization, with a booster immunization 14 days after the first inoculation. Mouse serum was collected 28 days after immunization, and the levels of HA1-specific antibodies in the mouse serum were detected by ELISA. (3) Detection of HA1 antibody in serum Antigen coating: HA1 (2 μg / ml) coating solution was added to Elisa plate (Corning 9018) at 100 ul / well and coated at 4°C overnight. Plate was washed once with 260 ul PBS per well. Blocked with 100 ul 5% blocking solution (5% FBS) at 37°C for 2 hours. Serum samples (1:10, 1:100, 1:1000, 1:10000, 1:100000…) were diluted in PBS and added to the blocked Elisa plate at 100 ul / well and incubated at 37°C for 1 hour. Washed 5 times with 260 ul PBST each time, 100 ul of enzyme-conjugated secondary antibody (enzyme-conjugated anti-mouse IgG-HRP 1:5000 diluted by PBS) was added to each well and incubated at 37°C for 1 hour. After washing twice with 260ul of PBST, 100ul / well of substrate TMB was added and incubated at room temperature for 15 minutes away from light to allow the substrate to develop color. 50ul of termination solution (2N H2SO4) was added to each well to terminate color development, and the plate was read with a microplate reader, with OD450-630. Titer calculation method: The maximum dilution factor that gave a positive result was selected, and the OD value / Cutoff value (0.1) corresponding to that dilution factor was multiplied by the dilution factor (X) to obtain the antibody titer corresponding to that serum. (4) 42 days after immunization, the mice were anesthetized and intranasally infected with 1,000 PFU of A / PR8 influenza virus. Starting from day 3 after virus infection, the mice were observed every two days and changes in body weight were recorded.

[0103] result: As shown in FIG. 26, after expressing and purifying the protein, the size and purity of the protein were detected by SDS-PAGE, and a single band was shown at the desired band size. As shown in FIG. 27, compared with HA1 protein, IFNα-HA1-Fc can induce higher titers of HA1-specific antibodies (FIG. 27a), indicating that the vaccine platform can enhance the immunogenicity of HA1 protein. After the mice are challenged, their body weight changes significantly, but the body weight of the mice in the IFNα-HA1-Fc immunized group can recover quickly compared with the PBS group and the HA1 protein immunized group. This indicates that immunization with IFNα-HA1-Fc vaccine has good protection against influenza infection (FIG. 27b).

[0104] Example 17 material and method: The design of IFNa-Pan-VZV-gE-Fc, IFNa-Pan-EBV-gp350-Fc, and IFNa-Pan-HSV-2-gD-Fc proteins, the construction of plasmids, and the purification of proteins were the same as in Examples 1 and 2.

[0105] result: As shown in Figure 28, after expression and purification of the IFNa-Pan-VZV-gE-Fc, IFNa-Pan-EBV-gp350-Fc, and IFNa-Pan-HSV-2-gD-Fc fusion proteins, the protein size and purity measurements by SDS-PAGE showed that the target bands were located at the correct positions.

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[18] MONTOYA M, SCHIAVONI G, MATTEI F, et al. Type I interferons produced by dendritic cells promote their phenotypic and functional activation [J]. Blood, 2002, 99(9): 3263-71.

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Claims

1. A vaccine comprising a fusion protein containing interferon-target antigen-immunoglobulin Fc region (or antibody Ab) as a structural unit, The interferon is a first structural unit and may be a type I interferon, a type II interferon and / or a type III interferon, such as IFN-α, IFN-β, IFN-γ, IFN-λ1 (IL-29), IFN-λ2 (IL-28a), IFN-λ (IL-28b) and IFN-ω, the interferon may be of human or murine origin, preferably the interferon is a type I interferon, such as IFN-α, such as mouse IFN-α4, human IFN-α2, a variant of human IFN-α2 (binding to human and mouse IFN receptors), such as those shown in the amino acid sequences SEQ ID NO: 1, SEQ ID NO: 21, SEQ ID NO: 22, The target antigen is a third structural unit, and the target antigen can be, for example, a tumor antigen, a pathogen antigen, for example, a viral or bacterial antigen, and the target antigen can be, for example, a mutant target antigen different from the wild type, and the mutant target antigen can include, for example, a natural point mutation / deletion mutation / gain mutation / truncation of the wild type antigen, an artificial point mutation / deletion mutation / gain mutation / truncation, any combination of natural or artificial mutation, and a subtype generated after mutation, and wherein the virus can be, for example, SARS-COV-2, or wherein the target antigen can be, for example, SARS-COV-2 virus S protein full length or S1 region, and for example, the target antigen can be the antigen shown in SEQ ID NO: 76 or SEQ ID NO: 77; The immunoglobulin Fc region (or antibody) is a second structural unit, and may be an amino acid sequence of a constant region of IgG1, IgG2, IgG3, IgG4, and / or IgM, such as an Fc region of IgG1, and an Fc region of IgG1-Fc-hole and IgG1-Fc-knob used to form a heterodimer, as shown in SEQ ID NO: 2, SEQ ID NO: 23, SEQ ID NO: 24, in which the antibody (including, for example, an antibody heavy chain or light chain, or a single-chain antibody; Ab for short) as a second structural unit may be a DC-targeting activating antibody, including antibodies such as anti-PD-L1, anti-DEC205, anti-CD80 / 86, etc. Optionally, said vaccine may be a targeted vaccine, and optionally said fusion protein may also comprise one or more Th cell helper epitopes and / or linking fragments.

2. The target antigen is a viral antigen, and the virus may be, for example, HBV, HPV, VZV, EBV, HSV-2, HIV, influenza virus, coronavirus, for example, SARS-COV, SARS-COV-2, MERS-CoV, and the antigen may be, for example, an HBV antigen, for example, an HBV Pres1 antigen, an HBsAg antigen, or a peptide, for example, an ad subtype or an ay subtype HBV Pres1 antigen, for example, an ad subtype HBV Pres1 antigen as shown in the amino acid sequence of SEQ ID NO: 6, for example, an ay subtype HBV Pres1 antigen as shown in the amino acid sequence of SEQ ID NO: 26; for example, an HBV HBsAg antigen (including each subtype and peptide), for example, an adr subtype HBV HBsAg antigen as shown in the amino acid sequence of SEQ ID NO: 7, for example, an adw subtype HBV HBsAg antigen as shown in the amino acid sequence of SEQ ID NO: 27, for example, an ay subtype HBV HBsAg antigen as shown in the amino acid sequence of SEQ ID NO:

28. For example, the antigen may be an HBsAg antigen; for example, the antigen may be, for example, a SARS-COV-2 antigen, for example, a SARS-COV2 RBD antigen; for example, a SARS-COV2 RBD antigen as shown in the amino acid sequence of SEQ ID NO: 8; for example, an influenza virus antigen, for example, an influenza virus HA antigen, for example, an influenza virus HA antigen as shown in the amino acid sequence of SEQ ID NO: 9; for example, an HPV antigen, for example, an HPV E7 antigen as shown in the amino acid sequence of SEQ ID NO: 10; for example, a gE antigen, for example, a varicella-zoster virus (VZV) gE antigen as shown in the amino acid sequence of SEQ ID NO: 91; for example, an EBV-gp350, for example, an Epstein-Barr virus (EBV) gp350 protein as shown in the amino acid sequence of SEQ ID NO: 92; for example, a gD antigen, for example, a Herpes Simplex Virus 2 (HSV-2) gD antigen as shown in the amino acid sequence of SEQ ID NO: 93; for example, an EBV It may be EBNA1 / LMP2, VZV-IE62, HSV-2 ICP0, HIV gp120 antigen; The target antigen may be a mutant viral antigen, for example, any mutant of the viral antigen, for example, a mutant of SARS-COV-2, for example, a naturally occurring point mutation / deletion mutation / gain mutation / truncated form, an artificial point mutation / deletion mutation / gain mutation / truncated form, any combination of natural or artificial mutations, or a subtype generated after mutation, of a SARS-COV-2 protein (e.g., at least one selected from the group consisting of S protein, N protein, M protein, and E protein). For example, the mutant viral antigen may be a full-length S protein (SEQ ID NO: 76), an S1 region (SEQ ID NO: 77), an RBD region (SEQ ID NO: 78), for example, the mutant viral antigen may include at least one mutation selected from the group consisting of a 69-70 deletion in the NTD region of the S protein of SARS-COV-2, a Y144 deletion, a 242-244 deletion, an L18F, D80A, D215, or R246I mutation, and a K417, E484, N501Y, or L452R mutation in the RBD region, a D614G, or H655Y mutation. For example, the mutant viral antigen may include a mutation derived from the UK B.1.1.7 (501Y.1) mutant strain, the South African B.1.351 (501Y.2) mutant strain, the Brazilian P1 (501Y.3) mutant strain, or the California B.1.429 mutant strain. For example, the mutant viral antigen may include a mutation derived from SEQ ID NO: 79, SEQ ID NO: 80, SEQ ID NO: 81, or SEQ ID NO: For example, the mutant viral antigen may comprise a mutant of a sequence as set forth in any one of SEQ ID NO: 79, SEQ ID NO: 80, SEQ ID NO: 81, SEQ ID NO: 82; The viral antigen may be fused with a helper polypeptide epitope that enhances B-cell and T-cell responses, and the helper polypeptide epitope may be located at the N-terminus or C-terminus of the antigen epitope, such as a Pan HLA DR-binding epitope (PADER), e.g., as shown in the amino acid sequence of SEQ ID NO: 3; The linked fragments of each structural unit are flexible polypeptide sequences, and can be linked fragments 1 and 2, e.g., those shown in SEQ ID NO: 4, SEQ ID NO: 25 amino acid sequences; the N-terminus of each polypeptide sequence of said structural units includes a corresponding signal peptide capable of promoting protein secretion, such as that shown in the amino acid sequence of SEQ ID NO: 5; The vaccine of claim 1, wherein the vaccine can be produced by a eukaryotic expression system, such as the eukaryotic expression system 293F, CHO cells.

3. The vaccine of claim 1 or 2, wherein the target antigen is a tumor antigen, for example a protein molecule highly expressed in tumor cells, for example, the antigen may be human epidermal growth factor receptor 2 (HER2 / neu) and epidermal growth factor receptor (EGFR); for example, the protein molecule highly expressed in tumor cells Her2 and each of its functional domains and truncations, for example, antigens shown in SEQ ID NOs: 85, 86, 97, 88, 89, 90 and mutants thereof.

4. the fusion protein is a homodimeric or heterodimeric fusion protein, and optionally the fusion protein may also comprise one or more Th cell helper epitopes and / or linking fragments in any one or two chains (i.e., the first polypeptide chain and / or the second polypeptide chain) of the homodimer or heterodimer; Optionally, the homodimeric fusion protein comprises a first polypeptide chain and a second polypeptide chain, the first polypeptide chain and the second polypeptide chain being completely identical, for example, the first polypeptide chain and the second polypeptide chain comprising, in N-terminal to C-terminal order, IFN, a target antigen and an immunoglobulin Fc region (or Ab), or forming a polypeptide in any combination order of the three structural units to generate a homodimer; preferably comprising, in N-terminal to C-terminal order, IFN, a target antigen and an immunoglobulin Fc region (or Ab); and may further comprise a fusion protein of a Th cell helper epitope; Optionally, the heterodimeric fusion protein comprises a first polypeptide chain and a second polypeptide chain, wherein the first polypeptide chain and the second polypeptide chain are not the same polypeptide chain, e.g., the first polypeptide chain may comprise, from N-terminus to C-terminus, an IFN and an immunoglobulin Fc region (or Ab), or may comprise, from N-terminus to C-terminus, an immunoglobulin Fc region (or Ab) and an IFN, and the second polypeptide chain may comprise a target antigen and an immunoglobulin Fc region (or Ab), wherein the target antigen is N-terminus The vaccine according to any one of claims 1 to 3, further comprising a fusion protein of a Th cell helper epitope, wherein the IFN and the target antigen are located at the N-terminus and the immunoglobulin region (or Ab) is located at the C-terminus, or the immunoglobulin region (or Ab) is located at the N-terminus and the target antigen is located at the C-terminus, or the polypeptide is formed in any combination order of the three structural units to generate a heterodimer; preferably, the IFN and the target antigen are located at the N-terminus of the two polypeptides, respectively, and the immunoglobulin Fc region (or Ab) is located at the C-terminus of the two polypeptides;

5. 1) the first and second polypeptides of the homodimer may comprise the amino acid sequences set forth in SEQ ID NOs: 11, 12, 13, 14, 29, 30, 31, 32, 38, 39, 40, 47, 48, 49, 50, 51, 56, 57, 59, 58, 65, 66, 67, 68; 2) the first polypeptide of the heterodimer may comprise a nucleotide sequence as set forth in SEQ ID NO: 15, 33, 42, 51, 60, 69, and the second polypeptide may comprise an amino acid sequence as set forth in SEQ ID NO: 16, 17, 18, 19, 34, 35, 36, 37, 43, 44, 45, 46, 52, 53, 54, 55, 61, 62, 63, 64, 70, 71, 72, 73; 3) The vaccine of claim 4, wherein the antibody comprises a DC targeting antibody, an immune checkpoint blocking antibody, an immune activating antibody, etc., such as an antibody amino acid sequence of anti-PD-L1 antibody (SEQ ID NO: 20), anti-DEC205 antibody, anti-CD80 / 86, etc.

6. A nucleic acid molecule encoding a fusion protein in a vaccine according to any one of claims 1 to 5, an expression vector comprising said nucleic acid molecule, or a host cell, such as a eukaryotic cell, comprising said nucleic acid molecule or expression vector.

7. Use of a fusion protein in a vaccine according to any one of claims 1 to 5 in the preparation of a composition, a kit, a recombinant microorganism or a cell line, said composition or said kit being for example a pharmaceutical composition, a pharmaceutical kit, an immunogenic composition or an immunogenic kit.

8. The use according to claim 7, wherein the composition or the kit is used for the prevention or treatment of a tumor or a pathogen, for example for the prevention or treatment of a virus or a bacterium, the virus may be HBV, HPV, EBV, influenza virus, HIV, coronavirus, such as SARS-COV, SARS-COV-2, MERS-CoV, for example, the composition or the kit is a hepatitis B prophylactic or therapeutic vaccine, an HBV prophylactic or therapeutic vaccine, an influenza prophylactic or therapeutic vaccine, a SARS-COV2 prophylactic or therapeutic vaccine, an HPV prophylactic or therapeutic vaccine, an HPV-associated tumor prophylactic or therapeutic vaccine, an EBV prophylactic or therapeutic vaccine, an EBV-associated tumor prophylactic or therapeutic vaccine, an HIV prophylactic or therapeutic vaccine.

9. The vaccine according to any one of claims 1 to 5, the use according to claim 7 or 8, wherein the vaccine, the composition or the kit can be administered by an immunization route such as intramuscular, intravenous, transdermal, subcutaneous or nasal, and the vaccine, the composition or the kit may further comprise an adjuvant, which may comprise aluminum adjuvant (Alum), Toll-like receptor 4 activator ligand MPLA, Toll-like receptor 9 ligand, oligodeoxynucleotide (CpG-ODN), M59, and Freund's adjuvant.

10. The vaccine may be used in combination with another prophylactic or therapeutic therapy, for example the vaccine is an HBV therapeutic vaccine, the HBV therapeutic vaccine may be used in combination with another prophylactic or therapeutic HBV therapy, for example the HBV therapeutic vaccine may be used in combination with a Hepatitis B virus envelope protein HBsAg vaccine, for example for the treatment of chronic Hepatitis B virus infection, for example the HBV therapeutic vaccine may be used in combination with a nucleoside or nucleotide analogue, for example for the treatment of chronic Hepatitis B virus infection, for example for use in combination with antiviral drugs and other therapies, such as influenza, SARS-COV2, HPV, EBV, HIV prophylactic or therapeutic vaccines, for use in combination with antiviral and tumor drugs and therapies, such as HPV, EBV related tumor prophylactic or therapeutic vaccines, for example a multivalent combination of any of the vaccines according to any one of claims 1 to 5 as a component of a vaccine and other viruses or pathogens or tumor vaccines. A vaccine, for example a multivalent combination vaccine consisting of any of the SARS-COV-2 vaccines according to any one of claims 1 to 5 and an influenza vaccine or other vaccine, for example any of the vaccines according to any one of claims 1 to 5, in a sequential or simultaneous immunization procedure with an adenoviral vaccine, mRNA vaccine, inactivated vaccine or DNA vaccine of the same virus, pathogen or tumor, for example a SARS-COV-2 fusion protein vaccine in a sequential or simultaneous immunization procedure with an adenoviral vaccine, mRNA vaccine, inactivated vaccine or DNA vaccine of SARS-COV-2, for example the order of immunization is 1) first immunization with the SARS-COV-2 fusion protein vaccine of the present invention and then immunization with another vaccine, 2) first immunization with an adenoviral vaccine, mRNA vaccine, inactivated vaccine or DNA vaccine of SARS-COV-2 and then immunization with a SARS-COV-2 fusion protein vaccine, 3) with a SARS-COV-2 fusion protein vaccine,The vaccine according to any one of claims 1 to 5, or the use according to claim 7 or 8, which is simultaneous immunization with an adenovirus vaccine, an mRNA vaccine, an inactivated vaccine or a DNA vaccine against SARS-COV-2.

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