A recombinant herpes zoster vaccine and its preparation method and application

Abstract

The present invention discloses a recombinant herpes zoster vaccine and its preparation method and application, and specifically relates to the field of biomedical technology. The recombinant herpes zoster vaccine includes a recombinant herpes zoster vaccine fusion protein purified product HN101 and a liposome mixture HA201, wherein the recombinant herpes zoster fusion protein gE-gD is reconstructed by reconstructing gE protein and reconstructing gD protein. The present invention reconstructs the gE protein of varicella-zoster virus and the gD protein of HSV through gene recombination technology, and then recombines to obtain the gE-gD recombinant herpes zoster fusion protein. After combining with the liposome mixture HA201, the gE-gD recombinant herpes zoster fusion protein of the present invention can induce a stronger immune response, improve antibody titer and cellular immunity, enhance vaccine effect, and avoid potential infection risks.

Description

Technical Field

[0001] The present invention relates to the field of biomedicine technology, and in particular to a recombinant herpes zoster vaccine and a preparation method and application thereof. Background Art

[0002] Herpes zoster (HZ) is caused by the varicella-zoster virus (VZV) that lurks in the body. Infection with the herpes zoster virus can activate acute infectious diseases. Infection with the herpes zoster virus can activate acute infectious diseases. VZV invades the human body through the upper respiratory tract or conjunctiva and causes systemic infection. The initial infection manifests as chickenpox in young children and can be latent infection in adults.

[0003] After recovery from the initial infection, VZV can remain dormant in the dorsal root ganglia of the spinal cord for a long time. When the body's immunity decreases, it can grow and reproduce again, causing HZ, which is usually manifested as unilateral skin herpes and pain distributed along the nerve segments. The most common complication of HZ is postherpetic neuralgia, which is the most common type of neuropathic pain. It can manifest as persistent pain or reappear after a period of relief, seriously affecting the patient's quality of life and bringing a heavy disease burden to society. According to statistics, about 1 / 4 of the world's population is at risk of developing HZ, of which 2 / 3 of HZ patients are 50 years old and above.

[0004] Although there are many methods to treat HZ, vaccines are still considered the most economical and effective measure to control HZ. However, the biggest drawback of vaccines is that their immunogenicity is often low and they cannot achieve good immune protection. They need to add adjuvants or delivery vectors to improve their immune efficacy.

[0005] Adjuvants are non-specific immunopotentiators that can enhance the body's immune response to antigens or change the type of immune response when injected with antigens or pre-injected into the body. Adjuvants can increase the biological or immunological half-life of vaccine antigens, enhance the delivery of antigens to antigen-presenting cells (APCs) and the processing and presentation of antigens by APCs, and induce the production of immunomodulatory factors. By regulating cytokine responses, adjuvant-containing vaccines can be designed to induce Th1 or Th2 responses. The advantages of using immune adjuvants in vaccine preparations are that they can enhance weaker immunogenicity (such as highly purified or recombinant antigens), reduce the amount of antigen or the number of vaccinations required to provide protective immunity, and improve the effectiveness of vaccines in those with weaker immune responses (such as newborns, the elderly, and immune-compromised vaccine recipients). However, due to the difficulty of research and development, the types of vaccine adjuvants that have been marketed worldwide are still scarce. At present, China still remains at the traditional level of aluminum adjuvants, which is the only immune adjuvant approved by the International Development Association (IDA) for use in humans and animals. This adjuvant has the function of antigen adsorption, and can adsorb protein antigens from the solution, and finally form antigen precipitation. After being injected into the body, it can slowly release the antigen, thereby prolonging the time of antigen action, and at the same time promoting the response of macrophages at the injection site. However, the biggest disadvantage of aluminum adjuvant is that the immune response produced is weaker than that of other applied adjuvants, and it cannot participate in cellular immune response.

[0006] Currently, the commercially available recombinant shingles vaccines in use are expensive, so there is an urgent need to solve the problems of the current shingles vaccines. Summary of the invention

[0007] To this end, the present invention provides a recombinant herpes zoster vaccine and a preparation method and application thereof to solve the above-mentioned problems.

[0008] In order to achieve the above object, the present invention provides the following technical solutions:

[0009] According to the first aspect of the present invention, a recombinant herpes zoster vaccine is provided, comprising a purified recombinant herpes zoster vaccine fusion protein HN101 and a liposome mixture HA201, wherein the recombinant herpes zoster fusion protein gE-gD is recombinantly constructed by reconstructing a reconstructed gE protein and a reconstructed gD protein.

[0010] In the recombinant herpes zoster vaccine, the concentration of the purified recombinant herpes zoster vaccine fusion protein HN101 is 0.08-0.12 mg / ml; as an example, 0.1 mg / ml is preferred.

[0011] HSV gD is a type I membrane glycoprotein, approximately 8-10 nm long, irregularly clustered on the surface of the viral membrane. HSV gD is organized into an extracellular domain, a TMD, and a short cytoplasmic tail. Although the gD of all alpha-herpesviruses has similar functions—binding to host cell receptors and initiating fusion reactions—it is irreplaceable. Experiments that have attempted to replace it have reported a complete loss of function.

[0012] According to crystallographic studies, the gD extracellular domain has an immunoglobulin-like core with N- and C-terminal extended edges at both ends. The N-terminal domain is called the receptor binding domain (RBD), and this part of gD binds to specific host receptors. The C-terminal domain is called the prefusion domain (PFD), and it interacts with gH / gL and gB. The PFD also binds to the N-terminal region and forms an auto-inhibitory closed conformation. This auto-inhibitory conformation is essential to prevent gD from binding to gH / gL or gB before binding to its specific receptor. The binding of gD to its specific receptor causes a conformational change in gD that favors the release of the PFD domain from its N-terminal interaction. Studies have shown that antibody binding to this specific region blocks the interaction of gH / gL and gB with gD.

[0013] Both the RBD and the PFD are essential for the fusion reaction. The infectivity of HSV-1 lacking gD can be restored upon addition of exogenous soluble gD, but only if the PFD and RBD regions of the gD ectodomain are present in this soluble form, as these regions cannot function independently. The RBD of gD is essential for its recognition of its receptors and for the binding of gB to its receptors, especially PILRα.

[0014] Therefore, after the HSV reconstructed gD and the VZV reconstructed gE are recombined again, the gE-gD recombinant herpes zoster fusion protein is obtained, which can induce a stronger immune response, increase antibody titers and cellular immunity, enhance vaccine effectiveness, and avoid potential infection risks.

[0015] Furthermore, the amino acid sequence of the recombinant herpes zoster fusion protein is shown in SEQ ID NO.1.

[0016] Furthermore, the gene encoding nucleotide sequence of the recombinant herpes zoster fusion protein is shown in SEQ ID NO.2.

[0017] Furthermore, the liposome mixture HA201 is an adjuvant system; it is also the key to the effectiveness of the vaccine and the bottleneck of domestic vaccines. The present invention obtains a new adjuvant system by adjusting the ratio and preparation method of the adjuvant system.

[0018] Furthermore, the preparation method of the liposome mixture HA201 includes: dissolving 3D-MLA, DOPC and cholesterol with isopropanol, evaporating and drying to form a liposome membrane, adding PBS buffer solution for hydration, homogenizing with a high-pressure homogenizer until the particle size reaches 90nm to 110nm, then adding QS-21, diluting, sterile filtering and then packaging.

[0019] According to the second aspect of the present invention, a method for preparing a recombinant herpes zoster fusion protein is provided, the method comprising:

[0020] Step 1, connecting the recombinant herpes zoster gE-gD fusion gene into an expression vector to construct an expression recombinant vector;

[0021] Step 2, transforming the constructed expression recombinant vector into a host cell to construct a recombinant gene host cell capable of expressing varicella-zoster virus gE-gD fusion protein;

[0022] Step three, using the recombinant gene host cell to express the recombinant herpes zoster virus gE-gD fusion protein and purifying it.

[0023] Furthermore, in step 1, the expression vector is pcDNA3.1(+) (Cat. No.: V79020).

[0024] Furthermore, in step 2, the host cell is a CHO cell; CHO cells (Chinese Hamster Ovary cells) are a laboratory-cultured cell line derived from Chinese hamster ovary cells.

[0025] Furthermore, in step three, purification includes ultrafiltration, low pH inactivation, multi-step purification and nanofiltration; this part of the main purification can also select conventional purification methods on the market such as hydrophobic chromatography, anion exchange chromatography, hydroxyapatite chromatography, ultrafiltration and nanofiltration.

[0026] According to the third aspect of the present invention, a recombinant herpes zoster fusion protein is provided for use in preparing a herpes zoster vaccine.

[0027] The present invention has the following advantages:

[0028] The present invention reconstructs the gE protein of varicella-zoster virus and the gD protein of HSV through gene recombination technology and then recombines them to obtain the gE-gD recombinant herpes zoster fusion protein. After combining with the liposome mixture HA201, the gE-gD recombinant herpes zoster fusion protein of the present invention can induce a stronger immune response, improve antibody titer and cellular immunity, enhance vaccine effect, and avoid potential infection risks.

[0029] The fusion protein prepared by the present invention can induce a strong immune response and can effectively prevent latent viral infection in subsequent animal experiments, thereby improving the safety of the vaccine and being a potential candidate to replace the existing attenuated live vaccine. BRIEF DESCRIPTION OF THE DRAWINGS

[0030] In order to more clearly illustrate the implementation methods of the present invention or the technical solutions in the prior art, the following briefly introduces the drawings required for the implementation methods or the description of the prior art. Obviously, the drawings in the following description are only exemplary, and for ordinary technicians in this field, other implementation drawings can be derived from the provided drawings without creative work.

[0031] The structures, proportions, sizes, etc. illustrated in this specification are only used to match the contents disclosed in the specification so as to facilitate understanding and reading by persons familiar with the technology. They are not used to limit the conditions under which the present invention can be implemented, and therefore have no substantial technical significance. Any structural modification, change in proportion or adjustment of size shall still fall within the scope of the technical contents disclosed in the present invention without affecting the effects and purposes that can be achieved by the present invention.

[0032] Figure 1 A simulated spatial conformation diagram of the herpes zoster virus gE-gD fusion protein provided in Example 1 of the present invention;

[0033] Figure 2 The test results of neutralizing antibody titer in mouse serum after vaccination with the vaccine of the present invention provided in Example 4 of the present invention;

[0034] Figure 3 The test results of lymphocyte proliferation effect on mice after vaccination with the vaccine of the present invention provided in Example 4 of the present invention;

[0035] Figure 4 This is a graph showing the results of detecting the levels of IFN-γ and IL-4 secreted by mouse lymphocytes after being stimulated by specific antigens after vaccination with the vaccine of the present invention as provided in Example 4 of the present invention. DETAILED DESCRIPTION

[0036] The following is a description of the implementation of the present invention by specific embodiments. People familiar with the art can easily understand other advantages and effects of the present invention from the contents disclosed in this specification. Obviously, the described embodiments are part of the embodiments of the present invention, not all of the embodiments. Based on the embodiments of the present invention, all other embodiments obtained by ordinary technicians in this field without creative work are within the scope of protection of the present invention.

[0037] Example 1

[0038] Recombinant construction of fusion protein gE-gD:

[0039] Through NCBI database and literature search, the amino acid sequence of gE protein extracellular region (NP_040190.1, 1-539aa) and a segment of effective gD protein (WPC89038.1, 54-269aa) were selected as the basis for gene optimization. After design, gE and gD were reconstructed respectively and then recombined. The two were used (GGGS) 3 After ligation, the purity of the purified fusion protein reached more than 95% by HPLC. The spatial structure of the recombinant protein was as follows Figure 1 As shown, the sequence of the recombinant herpes zoster gE-gD fusion protein is shown in SEQ ID NO.1.

[0040] SEQ ID NO.1:

[0041]

[0042] The recombinant herpes zoster virus gE-gD fusion protein is efficiently expressed in CHO cells. The codons preferred by CHO cells are selected to optimize the coding of the recombinant herpes zoster virus gE-gD gene (such as SEQ ID NO: 2), and the synthesis is entrusted to an outsourcing company, as follows. It should be noted that the optimization principle is not to simply select the codons with the highest frequency in CHO cells, but a more complex optimization scheme. There are three overall optimization principles: first, according to the degeneracy of the codons, the original codons are replaced with the high-frequency codons corresponding to each amino acid in the CHO cells; second, in order to avoid the excessive GC content in the transcribed mRNA affecting its secondary structure and thus affecting the translation efficiency, the GC% of the gene is controlled at about 50% during the optimization process; third, some commonly used restriction enzyme cutting sites are avoided.

[0043] SEQ ID NO 2:

[0044]

[0045]

[0046] Add 4×His tag, select pcDNA3.1(+) as vector, and perform codon optimization according to host CHO cells. Recombinate the gene sequence of the synthesized fusion protein into the target vector pcDNA3.1(+) to obtain pcDNA3.1(+)-gE-gD-4×His recombinant expression vector, and the reaction system is shown in Table 1.

[0047] Table 1

[0048] name Volume / μL Carrier 5 product 5 Assembly 10 Total volume 20

[0049] The above ligation solution was connected at a constant temperature of 52°C for 30 minutes to obtain the recombinant expression vector pcDNA3.1(+)-gE-gD-4×His.

[0050] Example 2

[0051] Expression and purification of the recombinant expression vector pcDNA3.1(+)-gE-gD-4×His:

[0052] The expression plasmid of Example 1 was prepared in large quantities and stably transfected into the host cell CHO-K1 after linearization. In this embodiment, a total of 10 transfection experiments were performed. Each group of transfected mini cell groups was then screened by batch feeding culture, and the cell groups with higher expression levels were selected for subsequent limited dilution cloning screening. It can be seen that in all 10 groups of mini cell groups, the average cell expression level was between 0.08-0.12 mg / ml, while the expression levels of the top three groups of cell groups were between 0.1-0.12 mg / ml, and the highest group of cell group protein expression levels could be 0.12 mg / ml. On this basis, the three groups of cell groups with the highest expression levels were selected (derived from different plasmids, but the cell growth conditions were all good), and monoclonal clones were selected from these three groups of cell groups by limiting dilution. The selected monoclones are expanded and cultured in the batch feeding process, the supernatants of the above cloned cells are collected, and samples are taken for western blot detection to determine the target protein according to the bands. The LDC photos, growth conditions, expression levels, live cell density, viability, terminal lactic acid content and related product quality parameters of the clones are comprehensively considered to select the best three clones, which are the dominant cell lines. The cell lines obtained above are cultured and expressed in a bioreactor to obtain the cell culture supernatant containing the recombinant gE-gD protein. The above supernatant can also be sampled for Western blot detection to determine whether it is the target protein according to the bands. It has been proven that the average value of gE protein in the above cell lines can reach 0.1 mg / ml; after conventional ultrafiltration, low pH inactivation, multi-step purification, and nanofiltration purification, the purity of the fusion protein is more than 95% detected by HPLC.

[0053] It should be noted that the method of using CHO cell lines to stably express gE-gD recombinant protein is a well-known method in the art, and specific references may be made to the Guidelines for Molecular Cloning Experiments and other public documents.

[0054] Example 3

[0055] Evaluation of the immunogenicity of recombinant herpes zoster vaccines:

[0056] The purified gE-gD protein solution with a purity of more than 95% obtained in Example 2 was mixed with the liposome mixture HA201 to obtain a recombinant herpes zoster vaccine; wherein the liposome mixture HA201: 40-60 mg of 3D-MLA, 800-1200 mg of DOPC and 200-300 mg of cholesterol were dissolved in 20-30 mL of isopropanol, evaporated and dried to form a liposome membrane, 40-60 ml of PBS buffer solution was added for hydration, homogenized by a high-pressure homogenizer until the particle size reached 90 nm-120 nm, and then 40-60 mg of QS-21 was added, and after dilution, the mixture was sterile filtered and packaged.

[0057] Carry out sterility test according to the current "Chinese Pharmacopoeia" and use the horseshoe crab reagent method for endotoxin detection. It can only be used if the endotoxin content is not higher than 100EU / mL.

[0058] Example 4

[0059] The vaccine of Example 3 was diluted to a gE-gD protein content of 60 μg / ml to prepare an animal experiment vaccine, with an injection volume of 0.5 mL / animal.

[0060] 1. Anti-infection of mice

[0061] C57BL / 6J mice were used as animal models for immunogenicity studies. Thirty C57BL / 6J mice aged 4-6 weeks were selected and randomly divided into 5 groups, with 6 mice in each group, including experimental group 1, commercial vaccine Shingrix control group 1, blank liposome mixture HA201 control group 2, blank control group 3 that was not injected with the vaccine but injected with the same dose of saline, and observation group without any treatment.

[0062] Treatment: Immunization was performed in the first and third weeks.

[0063] 2.ELISA antibody titer detection:

[0064] The recombinant VZV gE-gD protein was diluted to 1.5 μg / ml with carbonate buffer, coated on a 96-well ELISA plate (Thermo), 100 μl per well, and placed at 2-8°C overnight; the liquid in the 96-well plate was poured out, and the plate was washed 3 times with 20 mM PBST, and then 300 μl of blocking solution (5% skim milk powder) was added to each well, and the plate was blocked at room temperature for 1.5 hours; the blocking solution in the well was poured out, and the plate was washed 3 times with 20 mM PBS-T solution, and the mouse serum pre-diluted at 1:1000 (or 1:10000) was added to the first column of the 96-well ELISA plate, and then a 2-fold serial dilution was performed, and the negative control was the mouse serum subcutaneously immunized with PBS (1:1000 or 1:2000), and the plate was reacted at 37°C for 60 minutes, and the blocking solution in the well was poured out, and the plate was washed 3 times with 20 mM PBS-T solution. Wash 3 times with PBS-T solution; take out the goat anti-mouse IgG-HRP conjugate, dilute it with enzyme conjugate diluent at 1:20000, and then add it to a 96-well ELISA plate, 100μl per well, and react at 37℃ for 1 hour; aspirate the blocking solution in the wells, wash 5 to 6 times with 20mM PBS-T solution, add 50μl TMB colorimetric solution to each well, leave it at room temperature for 15 minutes, then add 50μl stop solution to terminate the reaction, and then use Shenzhen Huisong ELISA instrument to measure the A450 / A630 absorbance value, and use 2.1 times the A450 / A630 of the negative control group mixed serum as the Cut-Off value (if the negative control A450 value is lower than 0.05, calculate it as 0.05) to determine the titer of the immunized serum. The geometric mean and standard deviation of the anti-VZV gE-gD antibody titer in the serum of mice in each experimental group are shown in Table 2 below; after two immunizations, the serum antibodies of the mice in the experimental group and the mice in the comparison group 1 all showed positive conversion; statistical analysis of the antibody titers measured in each group of experimental animals showed that the ELISA titers of the mice in the experimental group and the mice in the comparison group 1 after two immunizations were between 400,000 and 1.44 million, and there was no significant difference in the ELISA titers between the mice in the experimental group and the commercially available group (comparison group 1).

[0065] Table 2

[0066]

[0067] Mouse serum was collected and inactivated at 56°C for 30 min. The serum was diluted 1:50 as the stock solution, and then diluted 1:2, 1:4, etc. to 1:128. 200 μL of the diluted serum was mixed with 200 μL of VZV virus with a titer of 5000 PFU / mL and incubated at 37°C for 1 hour. The mixture was then inoculated into a 24-well plate covered with a monolayer of MRC-5 cells and incubated at 37°C with 5% CO. 2 The cells were incubated in an incubator for 48 to 72 hours. The virus titer was calculated based on the number of plaques in the blank control (containing serum from non-immunized mice), and the neutralizing antibody titer (NT50) was determined based on the immune serum dilution that could inhibit 50% of the virus pathology. Figure 2As shown, the neutralizing antibody titers of the serum of mice immunized with the commercially available (control group 1) as antigen were 1:89 and 1:109 at 1 and 3 weeks after immunization, respectively; while the neutralizing antibody titers of the serum of mice immunized with the recombinant gE-gD fusion protein (experimental group) as antigen were 1:159 and 1:179 at 1 and 3 weeks after immunization, respectively; It can be seen that mice immunized with the recombinant 1 and 3 week fusion protein as antigen can produce higher titers of neutralizing antibodies.

[0068] Any antiserum with an ELISA titer of 1:1280000 was mixed, and then the neutralizing antibody titer was measured, and the result was 175.

[0069] 3. Cellular Immunity Detection

[0070] Three mice were killed at 1 and 3 weeks after immunization, and spleen lymphocytes were isolated aseptically. The total cell concentration was adjusted to 5 × 10 6 / mL. 100 μL of cell suspension was added to each well of a 96-well plate. Each mouse had 8 wells, and 1×10 4 PFU inactivated VZV was used as the test group, and 4 wells were added with culture medium as the negative control group. The cells were placed at 37°C and 5% CO 2 Culture in an incubator for 48 hours. Take 100 μL of culture supernatant from each well of the 7th week mice to measure the IFN-γ and IL-4 content. At the same time, add 20 μL MTT (5 mg / mL) to each well and continue to culture for 4 hours. Discard the culture supernatant, add 100 μL DMSO to each well, dissolve the crystals, and read the OD 570 Value. OD of experimental group 570 OD 570 The ratio between the values ​​is the stimulation index (SI). The larger the stimulation index, the stronger the lymphocyte proliferation ability. Figure 3 As shown in the figure, the lymphocyte stimulation index of mice immunized with the commercial vaccine (control group 1) as antigen was 2.4 and 2.5 at 1 and 3 weeks after immunization, respectively; while the lymphocyte stimulation index of mice immunized with the recombinant gE-gDL fusion protein (experimental group) as antigen was 3.9 and 3.5 at 1 and 3 weeks after immunization, respectively. Therefore, mice immunized with the recombinant gE-gD fusion protein (experimental group) as antigen can induce stronger cellular immunity than commercial vaccines. After the mouse lymphocytes were stimulated with inactivated VZV in vitro at week 3 after immunization, as shown in the figure, Figure 4 As shown, the average IFN-γ content in the cell culture supernatant of the control group 1 was 125.9pg / mL, and the recombinant gE-gDL fusion protein group (experimental group) was 187pg / mL, both higher than the observation group (35.7pg / mL); and the concentration of IL-4 in the immunization group was about 2 times higher than that in the control group, and there was no significant difference between the different antigen immunization groups. These indicate that vaccination with this subunit vaccine may form a strong Th1 type cellular immune memory.

[0071] 4. Virus attack and protection experiment

[0072] A VZV strain that can adapt to guinea pig cells is cultured in vitro, and then the strain is used to infect guinea pig peripheral blood lymphocytes in vitro. The guinea pig lymphocytes infected with VZV are then transfused back into the guinea pigs. After 28 days, latent infection can be established in the intestinal ganglia and dorsal root ganglia of the guinea pigs. This model was used to conduct a virus attack and protection test. Fifteen 10-week-old female FMMU albino guinea pigs were divided into three groups, with 5 in each group. The first group was the immunization group, which was immunized twice with the recombinant herpes zoster vaccine prepared by the present invention (the second immunization was performed 14 days after the first immunization, 30 μg per pig each time, subcutaneously injected). The second group was the saline control group, which was injected with the same volume of sterile saline. The third group was the commercially available vaccine Shingrix. 28 days after the second immunization, guinea pig PBMC infected with VZV were prepared. The steps are as follows: MRC-5 cells were added to a 6-well plate and cultured until the cells grew into a monolayer, and 5×10 5 PFU VZV was cultured for 24 h and 3 × 10 6 guinea pig PBMCs were centrifuged at 200 × g for 45 min at room temperature and the 6-well plate was placed at 33°C with 5% CO 2 Continue to culture in the incubator for 20 hours, gently blow up the PBMC, centrifuge at 420×g for 5 min at room temperature, discard the supernatant, resuspend the cells with fresh saline, and adjust the cell concentration to 3×10 6 / 50μL, 50μL of guinea pig PBMC infected with VZV was returned to the guinea pig blood circulation through the ophthalmic venous sinus, and the guinea pigs were killed 28 days later. The enteric ganglia and dorsal root ganglia were separated from the intestinal tissue and spine, and DNA was extracted. VZV ORF29 and ORF40 genes were detected by nested PCR. As long as one of the genes was positive, it was considered that VZV infection existed, and both were negative, which meant that there was no VZV infection. The results are shown in Table 3. VZV DNA was not detected in the ganglia of guinea pigs after vaccination, while VZV DNA was detected in guinea pigs that were not vaccinated, indicating that the vaccine has a good protective effect.

[0073] Table 3

[0074]

[0075] Although the present invention has been described in detail above by general description and specific embodiments, it is obvious to those skilled in the art that some modifications or improvements can be made to the present invention. Therefore, these modifications or improvements made without departing from the spirit of the present invention all belong to the scope of protection claimed by the present invention.

Claims

1. A recombinant herpes zoster vaccine, characterized in that: It includes a purified recombinant herpes zoster vaccine fusion protein HN101 and a liposome mixture HA201, wherein the recombinant herpes zoster fusion protein gE-gD is reconstructed by recombining a reconstructed gE protein and a reconstructed gD protein; The amino acid sequence of the purified recombinant herpes zoster fusion protein HN101 is shown in SEQ ID NO.1; The gene encoding nucleotide sequence of the purified recombinant herpes zoster fusion protein HN101 is shown in SEQ ID NO.2; The liposome mixture HA201 is an adjuvant system with a particle size range of 90 to 110 nm. The preparation method of the liposome mixture HA201 comprises: dissolving 3D-MLA, DOPC and cholesterol with isopropanol, evaporating and drying to form a liposome membrane, adding PBS buffer solution for hydration, homogenizing with a high-pressure homogenizer until the particle size reaches 90 nm to 110 nm, adding QS-21, diluting, sterile filtering and then packaging.

2. A method for preparing a recombinant herpes zoster fusion protein, characterized in that: The method comprises: Step 1, connecting the recombinant herpes zoster gE-gD fusion gene into an expression vector to construct an expression recombinant vector; Step 2, transforming the constructed expression recombinant vector into a host cell to construct a recombinant gene host cell capable of expressing herpes zoster virus gE-gD fusion protein; the host cell is a CHO cell; Step 3, using the recombinant gene host cell to express the recombinant herpes zoster virus gE-gD fusion protein, and purifying it to obtain the purified recombinant herpes zoster fusion protein HN101; The amino acid sequence of the purified recombinant herpes zoster fusion protein HN101 is shown in SEQ ID NO.1; The gene encoding nucleotide sequence of the purified recombinant herpes zoster fusion protein HN101 is shown in SEQ ID NO.

2.

3. The method for preparing a recombinant herpes zoster fusion protein according to claim 2, characterized in that: In the step 1, the expression vector is pcDNA3.1(+).

4. The method for preparing a recombinant herpes zoster fusion protein according to claim 2, characterized in that: In the step 3, purification includes ultrafiltration, low pH inactivation, multi-step purification and nanofiltration.

5. Use of the purified recombinant herpes zoster fusion protein HN101 prepared by any of the methods of claims 2 to 4 in the preparation of a herpes zoster vaccine.

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