WO2021164097A1 - 用于预防新型冠状病毒的生物制品 - Google Patents

用于预防新型冠状病毒的生物制品 Download PDF

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WO2021164097A1
WO2021164097A1 PCT/CN2020/080861 CN2020080861W WO2021164097A1 WO 2021164097 A1 WO2021164097 A1 WO 2021164097A1 CN 2020080861 W CN2020080861 W CN 2020080861W WO 2021164097 A1 WO2021164097 A1 WO 2021164097A1
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protein
novel coronavirus
gene
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protein gene
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张书元
徐卫
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赛诺（深圳）生物医药研究有限公司
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    • A61K48/005—Medicinal preparations containing genetic material which is inserted into cells of the living body to treat genetic diseases; Gene therapy characterised by an aspect of the 'active' part of the composition delivered, i.e. the nucleic acid delivered
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    • A61P31/00—Antiinfectives, i.e. antibiotics, antiseptics, chemotherapeutics
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    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
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    • A61K39/00—Medicinal preparations containing antigens or antibodies
    • A61K2039/51—Medicinal preparations containing antigens or antibodies comprising whole cells, viruses or DNA/RNA
    • A61K2039/53—DNA (RNA) vaccination
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    • C12N2770/20011—Coronaviridae
    • C12N2770/20034—Use of virus or viral component as vaccine, e.g. live-attenuated or inactivated virus, VLP, viral protein

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  • the invention belongs to the technical field of biopharmaceutical products, and relates to biological products for the prevention of new coronaviruses. It uses gene synthesis, codon optimization, and gene cloning to construct recombinant adenovirus vector gene vaccines, and is used for the development of prevention of new coronaviruses (COVID-19). Gene vaccine products.
  • virus vaccine development methods include 1) inactivated virus vaccines, 2) attenuated virus vaccines, 3) subunit vaccines, 4) VLP (virus like particle) vaccines, 5) DNA plasmid vaccines, and 6) recombinant virus vector vaccines .
  • the use of high-tech recombinant viral vector gene vaccine is the latest technology in vaccine research and development. The vaccine is safe and effective, and the research and development time is short. Recently, on December 19, 2019, the US FDA approved a vaccine product (Merck, Ervebo, USA) for the prevention of Ebola virus infection, which is a recombinant viral vector gene vaccine (rVSV-ZEBOV) product.
  • vaccines using recombinant adenovirus vectors have achieved good clinical effects in the treatment and prevention of MERS coronavirus infections similar to the new coronavirus. It provides a scientific basis for the use of recombinant adenovirus vectors to develop new coronavirus vaccines.
  • the recombinant adenovirus vector vaccine has some outstanding advantages, including:
  • Adenovirus vectors are the most widely used viral vectors, and their safety is widely recognized.
  • the clinical data of the recombinant adenovirus products that have been on the market for more than ten years can provide sufficient evidence;
  • Vaccine products are suitable for large-scale GMP production and a wide range of public preventive treatment applications;
  • the vaccine product has high stability and can survive for more than 10 years at -80°C, which is suitable for strategic storage and standby.
  • Recombinant viral vector gene vaccine is one of the latest vaccine development technologies recognized globally. At present, the prevalence of the next-generation rapid gene sequence detection technology ensures that scientists can classify and accurately sequence the gene sequences of disease-causing viruses in a relatively short period of time. Using precise gene cloning and modification technology, a recombinant viral vector gene vaccine expressing the main antigen of the virus can be constructed at the fastest speed and accurately. The expressed viral antigen maintains the structure and framework of the wild virus, ensuring that a strong preventive immune response can be triggered in the patient's body. The preparation of traditional attenuated or inactivated vaccines requires live virus strains.
  • the process of attenuation or inactivation will also affect the integrity of important viral antigens, thereby affecting the ability to trigger an immune response in patients receiving the vaccine.
  • the vaccine construction process does not involve live virus strains, ensuring the safety of the construction process.
  • the new coronavirus is an enveloped positive-strand RNA virus with a genome length of 29.9Kbp.
  • Figure 1 shows a schematic diagram of the structure of the novel coronavirus particle and the main viral structural proteins.
  • S1 Spike S1 subunit
  • N Nucleocapsid
  • the spike protein (Spike, S protein) on the surface of the new coronavirus first interacts with the receptors of the infected patient's cells and attaches to the cell surface.
  • S protein is composed of two subunits, S1 and S2 subunits.
  • the S1 subunit includes the function of the receptor binding domain of the virus, and the S2 subunit includes the function of fusing the viral membrane and the cell membrane.
  • Peptidase on the cell surface is one of the main viral receptors.
  • the virus After the virus attaches to the cell surface, it undergoes acid-dependent proteolysis by cathepsin to cut the enzyme cut site between the S1 and S2 subunits, cleaves the S protein, initiates the fusion of the viral membrane and the cell membrane, and transports the viral genome into the cell. , Causing infection.
  • the S1 subunit of the extracellular domain of the spike protein is the main part that interacts with the cell surface receptor.
  • the virus's nucleocapsid N protein is the main structural protein of the virus. According to the structure of the new coronavirus and the mechanism of infecting patients, the S1 (spike) protein and N protein of the virus are the main viral antigens that trigger a strong immune response and are the best viral antigens for the development of vaccines.
  • the purpose of the present invention is to construct a biological product for preventing the novel coronavirus (COVID-19).
  • the biological product can be a genetic vaccine or a genetic medicine.
  • the following takes the construction of a recombinant adenovirus vector genetic vaccine as an example.
  • the recombinant adenovirus vector gene vaccine used to prevent the new coronavirus is constructed by using the expression vector tower to carry the antigen protein gene expression cassette of the new coronavirus.
  • the in vivo expression of the antigen protein gene activates the body's immune response to the new coronavirus to achieve prevention Novel coronavirus infection and transmission.
  • the structure of the antigen protein gene expression cassette includes the CMV promoter, the Kozak sequence, the antigen protein gene and the SV40polyA sequence connected in sequence, that is, the structure can be expressed as "CMV promoter-Kozak sequence-antigen protein gene-SV40polyA sequence.
  • the antigen protein gene is the S1 protein gene expressing the S1 protein antigen, or the S1 protein gene-T2A-N protein gene that simultaneously expresses the S1 protein antigen and the N protein antigen.
  • the codons are optimized first, and the gene fragments of the optimized S1 protein gene and the N protein gene are synthesized using conventional gene synthesis methods.
  • the S1 protein gene and/or the N protein gene are artificially synthesized and codon-optimized gene sequences of the novel coronavirus.
  • the nucleotide sequence of the codon-optimized S1 protein gene is as shown in SEQ ID NO.1 Show.
  • the nucleotide sequence of the codon-optimized N protein gene is shown in SEQ ID NO.2.
  • the present invention constructs an S1 protein gene expression cassette expressing S1 protein to achieve high expression of S1 protein.
  • Figure 2 shows the structure of the S1 protein gene expression cassette.
  • the nucleotide sequence of the S1 protein gene expression cassette is shown in SEQ ID NO.3.
  • the present invention also constructs an S1 protein and N protein gene expression cassette that simultaneously expresses the S1 protein and the N protein, so as to achieve the simultaneous high expression of the S1 protein and the N protein.
  • Figure 3 shows the structure of the S1 protein and N protein gene expression cassettes.
  • S1 protein and N protein gene expression cassettes are shown in SEQ ID NO.4.
  • the constructed gene expression cassettes were cloned into the expression shuttle plasmid of pAdEasy system.
  • the human type 5 adenovirus plasmid expressing the shuttle plasmid and pAdEasy system with the E1 and E3 genes deleted were simultaneously transduced into HEK293 cells.
  • recombinant adenovirus vector gene vaccine seeds expressing S1 protein antigen and simultaneously expressing S1 and N protein antigens are respectively produced.
  • DNA sequencing confirmed that the vaccine seeds carry the correct S1 protein and N protein gene sequences and the correct gene structure. It is further expanded in HEK293 cells through cell culture to establish a vaccine seed bank for large-scale production and clinical applications.
  • Figure 1 is a schematic diagram of the structure of a novel coronavirus particle.
  • FIG. 2 is a schematic diagram of the structure of the S1 protein gene expression cassette.
  • FIG. 3 is a schematic diagram of the structure of the S1 protein and N protein gene expression cassettes.
  • the S1 protein gene expression cassette uses the CMV promoter and adds the Kozak sequence. SV40 polyA sequence is used to ensure correct protein expression. Using conventional molecular biology methods and reagents, a complete S1 protein gene expression cassette was cloned and constructed. A schematic diagram of the S1 protein gene expression cassette is shown in FIG. 2.
  • the nucleotide sequence of the S1 protein gene expression cassette is shown in SEQ ID NO. 3, and the length is 2703 bp.
  • the constructed S1 protein gene expression cassette was cloned into the expression shuttle plasmid of the commercial pAdEasy system according to the method provided by the supplier.
  • the human type 5 adenovirus plasmid expressing the shuttle plasmid and pAdEasy system with the E1 and E3 genes deleted were simultaneously transduced into HEK293 cells. Through homologous recombination of genes in HEK293 cells, recombinant adenovirus vector gene vaccine seeds expressing S1 protein antigen are produced.
  • the constructed vaccine seed can be further expanded in HEK293 cells through cell culture to establish a vaccine seed bank for the production and clinical application of large-scale vaccine products.
  • Example 2 Construction of recombinant adenovirus vector gene vaccine seeds expressing S1 protein antigen and N protein antigen at the same time
  • S1 protein gene sequence and N protein gene sequence (GenBank: MN908947.3) of the new coronavirus to increase the protein expression rate in the human body.
  • the optimized S1 nucleotide sequence is shown in SEQ ID NO.1
  • the N nucleotide sequence is shown in SEQ ID NO.2.
  • a gene expressing T2A cleavable polypeptide was added between the S1 protein gene and the N protein gene to form a composite gene fragment expressing S1 protein and N protein.
  • the gene fragments expressing S1 protein and N protein were synthesized by ordering genes from IDT Corporation of the United States. Gene sequencing verified the correctness of the gene sequence.
  • the S1 protein and N protein gene expression cassette adopts the CMV promoter and adds the Kozak sequence.
  • SV40 polyA sequence is used to ensure correct protein expression.
  • clone and construct a complete S1 protein and N protein compound gene expression cassette, that is, the S1 protein and N protein gene expression cassette is shown in FIG. 3.
  • the nucleotide sequence of the S1 protein and N protein gene expression cassettes is shown in SEQ ID NO. 4, and the length is 4032 bp.
  • the constructed composite gene expression cassette of S1 protein and N protein was cloned into the expression shuttle plasmid of the commercial pAdEasy system according to the method provided by the supplier.
  • the human type 5 adenovirus plasmid expressing the shuttle plasmid and pAdEasy system with the E1 and E3 genes deleted were simultaneously transduced into HEK293 cells.
  • recombinant adenovirus vector gene vaccine seeds that can express S1 protein antigen and N protein antigen at the same time are produced.
  • DNA sequencing confirmed that the vaccine seed carries the correct S1 protein and N protein composite gene expression cassette sequence, and the nucleotide sequence is as shown in SEQ ID NO.4.
  • the constructed vaccine seed can be further expanded in HEK293 cells through cell culture to establish a vaccine seed bank for the production and clinical application of large-scale vaccine products.

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Abstract

提供了一种用于预防新型冠状病毒(COVID-19)的生物制品。所述生物制品可以为基因疫苗或基因药物，其中，基因疫苗采用E1和E3基因缺失的人5型腺病毒作为载体，搭载表达新型冠状病毒的Spike S1亚单位的S1蛋白抗原或同时搭载表达S1蛋白抗原和N蛋白抗原，可产生免疫反应，预防新型冠状病毒感染和传染。

Description

用于预防新型冠状病毒的生物制品 技术领域
本发明属于生物制药制品技术领域，涉及到预防新型冠状病毒的生物制品，利用基因合成，密码子优化，基因克隆构建重组腺病毒载体基因疫苗，用于开发预防新型冠状病毒(COVID-19)的基因疫苗产品。
背景技术
新型冠状病毒(COVID-19)的突发给社会公共健康带来了巨大的危机，感染传染性非常强，给人类造成了巨大的社会健康和经济影响，受到人们的高度重视，目前还没有有效的治疗方法和预防疫苗。为了人民大众的健康，国家和社会的安全，急需开发可以预防新型冠状病毒感染的疫苗。
常用的病毒疫苗研发方法包括1)灭活病毒疫苗，2)减活病毒疫苗，3)亚单位疫苗，4)VLP(virus like particle)疫苗，5)DNA质粒疫苗，和6)重组病毒载体疫苗。采用高科技重组病毒载体基因疫苗是疫苗研发的最新技术，疫苗安全有效，研发时间短。美国FDA在最近，2019年12月19日，批准上市的用于预防埃博拉(Ebola)病毒感染的疫苗产品(美国Merck公司，Ervebo)，就是一款重组病毒载体基因疫苗(rVSV-ZEBOV)产品。
其中采用重组腺病毒载体的疫苗在治疗预防和新型冠状病毒类似的MERS冠状病毒感染已取得了良好的临床效果。为采用重组腺病毒载体研发新型冠状病毒疫苗提供了科学依据。除此之外，重组腺病毒载体疫苗还有一些突出的优点，包括：
1)疫苗安全性高。腺病毒载体为运用最为广泛的病毒载体，安全性受到广泛认可。已上市重组腺病毒产品“今又生”十余年的临床数据均能给予充分的证明；
2)接种后疫苗抗原蛋白表达速度快，表达量高。可以快速的，有效的引发抗新型冠状病毒的预防免疫；
3)疫苗研发速度快，技术成熟。在有了新型冠状病毒RNA基因序列后，可以快速的，在3-4个星期内，完成构建疫苗种子；
4)现成的疫苗生产技术；
5)疫苗产品适合大规模GMP生产，和广泛的大众预防治疗应用；
6)疫苗产品稳定性高，在-80℃温度下可存活10年以上，适合战略储存，备用。
重组病毒载体基因疫苗是全球公认的最新的疫苗开发技术之一。目前，下一代快速基因序列检测技术的普遍，保证了科学人员可以在较短的时间内能分类和准确测序引发疾病病毒的基因序列。采用精确的基因克隆和修饰技术，可以以最快的速度，准确的构建表达病毒主要抗原的重组病毒载体基因疫苗。所表达的病毒抗原保持了野性病毒的结构和构架，确保在患者体内可以引发强有力的预防免疫反应。传统的减活或灭活疫苗的制备，需要活病毒株，减活或灭活过程也会影响到重要病毒抗原的完整性，从而会影响在接受接种疫苗患者体内引发免疫反应的能力。另外，疫苗构建过程不涉及到活病毒株，保证了构建过程的安全性。
发明内容
新型冠状病毒是包膜正链RNA病毒，基因组长度为29.9Kbp。在图1中显示了新型冠状病毒颗粒的结构示意图和主要病毒结构蛋白。
根据新型冠状病毒感染患者的机制，经研究发现，病毒的S1(Spike S1亚单位)蛋白和N(Nucleocapsid)蛋白具有很强的免疫性，可以在患者体内引发抗体和T细胞反应，是研发疫苗的最佳病毒抗原。
为了感染患者，新型冠状病毒表面的刺突蛋白(Spike,S蛋白)首先和感染患者细胞的受体相互作用，附着在细胞表面。S蛋白有两个亚单位组成，S1和S2亚单位。S1亚单位包括病毒的受体结合域功能，S2亚单位包括融合病毒膜和细胞膜的功能。细胞表面的肽酶是主要的病毒受体之一。在病毒附着在细胞表面后，通过组织蛋白酶，进行酸依赖性蛋白水解切割位于S1和S2亚单位之间的酶切点，切割S蛋白，引发融合病毒膜和细胞膜，将病毒基因组转运到细胞内，造成感染。
其中刺突蛋白胞外域的S1亚单位是和细胞表面受体作用的主要部分，除了刺突蛋白外，病毒的核壳N蛋白(nucleocapsid)是病毒的主要结构蛋白。根据新型冠状病毒的结构和感染患者的机制，病毒的S1(spike)蛋白和N蛋白是引发强免疫反应的主要病毒抗原，是研发疫苗的最佳病毒抗原。
本发明的目的是构建一种用于预防新型冠状病毒(COVID-19)的生物制品，该生物制品可以为基因疫苗或基因药物，如下以构建重组腺病毒载体基因疫苗为例。
用于预防新型冠状病毒重组腺病毒载体基因疫苗，是利用表达载体塔载新型冠状病毒的抗原蛋白基因表达盒构建，藉由抗原蛋白基因的体内表达激活肌体对新型冠状病毒 的免疫反应，达到预防新型冠状病毒感染和传染。
抗原蛋白基因表达盒的结构包括依次连接的CMV启动子、Kozak序列、抗原蛋白基因和SV40polyA序列，即结构可表示为“CMV启动子-Kozak序列-抗原蛋白基因-SV40polyA序列。
抗原蛋白基因为表达S1蛋白抗原的S1蛋白基因，或同时表达S1蛋白抗原和N蛋白抗原的S1蛋白基因-T2A-N蛋白基因。
按照公开发表的新型冠状病毒的基因序列(GenBank:MN908947.3)，首先进行密码子优化，采用常规的基因合成方法，基因合成优化后的S1蛋白基因和N蛋白基因的基因片段。
其中，S1蛋白基因和/或N蛋白基因为人工合成且经密码子优化后的新型冠状病毒的基因序列，优选地，密码子优化的S1蛋白基因的核苷酸序列如SEQ ID NO.1所示。密码子优化的N蛋白基因的核苷酸序列如SEQ ID NO.2所示。
采用常规的基因克隆方法，本发明构建了表达S1蛋白的S1蛋白基因表达盒，来达到S1蛋白的高度表达，在图2中显示了S1蛋白基因表达盒的结构。
S1蛋白基因表达盒的核苷酸序列如SEQ ID NO.3所示。
为了最大程度的提高免疫反应，本发明也构建了同时表达S1蛋白和N蛋白的S1蛋白和N蛋白基因表达盒，来达到S1蛋白和N蛋白的同时高度表达。在图3中显示了S1蛋白和N蛋白基因表达盒的结构。
S1蛋白和N蛋白基因表达盒的核苷酸序列如SEQ ID NO.4所示。
将构建的上述基因表达盒分别克隆到pAdEasy系统的表达穿梭质粒中。将表达穿梭质粒和pAdEasy系统的删除了E1和E3基因的人5型腺病毒质粒同时转导到HEK293细胞内。通过在HEK293细胞内进行基因同源重组，分别产生表达S1蛋白抗原和同时表达S1和N蛋白抗原的重组腺病毒载体基因疫苗种子。
通过DNA测序确认疫苗种子携带正确的S1蛋白和N蛋白基因序列和正确的基因结构。通过细胞培养在HEK293细胞内进一步扩增，建立疫苗种子库，用于大规模生产和临床应用。
附图说明
图1为新型冠状病毒颗粒的结构示意图。
图2为S1蛋白基因表达盒的结构示意图。
图3为S1蛋白和N蛋白基因表达盒的结构示意图。
具体实施方式
以下实施例用于说明本发明，但不用来限制本发明的范围。
若未特别指明，实施例中所用的技术手段为本领域技术人员所熟知的常规手段，实施例中所用的生化试剂、材料均为市售可得。
实施例1 表达S1蛋白抗原的重组腺病毒载体基因疫苗种子的构建
使用IDT公司的密码子优化软件，对新型冠状病毒的S1蛋白基因序列(GenBank:MN908947.3)进行优化，以提高在人体内的蛋白表达率。优化后的核苷酸序列如SEQ ID NO.1所示。从美国IDT公司订购基因合成了S1蛋白的基因片段。基因测序验证了基因序列的正确性。
为了实现快速和高量地表达S1蛋白，S1蛋白基因表达盒采用CMV启动子并添加了Kozak序列。采用SV40 polyA序列来保证正确的蛋白质的表达。采用常规的分子生物学方法和试剂，克隆组建完整的S1蛋白基因表达盒。在图2中显示了S1蛋白基因表达盒的示意图。
S1蛋白基因表达盒的核苷酸序列如SEQ ID NO.3所示，长度为2703bp。
将构建的S1蛋白基因表达盒，按照供应商提供的方法，克隆到已商业化的pAdEasy系统的表达穿梭质粒中。将表达穿梭质粒和pAdEasy系统的删除了E1和E3基因的人5型腺病毒质粒同时转导到HEK293细胞内。通过在HEK293细胞内进行基因同源重组，产生表达S1蛋白抗原的重组腺病毒载体基因疫苗种子。
通过DNA测序确认了疫苗种子携带正确的S1蛋白基因表达盒序列，核苷酸序列正如SEQ ID NO.3所示。
所构建的疫苗种子，可以通过细胞培养在HEK293细胞内进一步扩增，建立疫苗种子库，用于大规模疫苗产品的生产和临床应用。
实施例2 同时表达S1蛋白抗原和N蛋白抗原的重组腺病毒载体基因疫苗种子的构建
使用IDT公司的密码子优化软件，对新型冠状病毒的S1蛋白基因序列和N蛋白基因序列(GenBank:MN908947.3)进行优化，以提高在人体内的蛋白表达率。优化后的S1核苷酸序列如SEQ ID NO.1所示，N核苷酸序列如SEQ ID NO.2所示。为了实现同时等 当量的表达S1蛋白抗原和N蛋白抗原，在S1蛋白基因和N蛋白基因之间端添加了表达T2A可裂解的多肽的基因，构成了表达S1蛋白和N蛋白的复合基因片段。从美国IDT公司订购基因合成了表达S1蛋白和N蛋白的基因片段。基因测序验证了基因序列的正确性。
为了实现快速和高量地同时表达S1蛋白和N蛋白，S1蛋白和N蛋白基因表达盒采用了CMV启动子并添加了Kozak序列。采用SV40 polyA序列来保证正确的蛋白质的表达。采用常规的分子生物学方法和试剂，克隆组建完整的S1蛋白和N蛋白的复合基因表达盒，即S1蛋白和N蛋白基因表达盒。在图3中显示了S1蛋白和N蛋白基因表达盒基因表达盒的示意图。
S1蛋白和N蛋白基因表达盒的核苷酸序列如SEQ ID NO.4所示，长度为4032bp。
将构建的S1蛋白和N蛋白的复合基因表达盒，按照供应商提供的方法，克隆到已商业化的pAdEasy系统的表达穿梭质粒中。将表达穿梭质粒和pAdEasy系统的删除了E1和E3基因的人5型腺病毒质粒同时转导到HEK293细胞内。通过在HEK293细胞内进行基因同源重组，产生可以同时表达S1蛋白抗原和N蛋白抗原的重组腺病毒载体基因疫苗种子。
通过DNA测序确认了疫苗种子携带正确的S1蛋白和N蛋白的复合基因表达盒序列，核苷酸序列正如SEQ ID NO.4所示。
所构建的疫苗种子，可以通过细胞培养在HEK293细胞内进一步扩增，建立疫苗种子库，用于大规模疫苗产品的生产和临床应用。
虽然，上文中已经用一般性说明及具体实施方案对本发明作了详尽的描述，但在本发明基础上，可以对之作一些修改或改进，这对本领域技术人员而言是显而易见的。因此，在不偏离本发明精神的基础上所做的这些修改或改进，均属于本发明要求保护的范围。

Claims (10)

  1. 用于预防新型冠状病毒的生物制品，其特征在于，利用表达载体塔载新型冠状病毒的抗原蛋白基因表达盒构建，藉由抗原蛋白基因的体内表达激活肌体对新型冠状病毒的免疫反应，达到预防新型冠状病毒感染和传染。
  2. 如权利要求1所述的用于预防新型冠状病毒的的生物制品，其特征在于，所述抗原蛋白基因表达盒的结构包括依次连接的CMV启动子、Kozak序列、抗原蛋白基因和SV40polyA序列。
  3. 如权利要求2所述的用于预防新型冠状病毒的的生物制品，其特征在于，所述抗原蛋白基因为表达S1蛋白抗原的S1蛋白基因，或同时表达S1蛋白抗原和N蛋白抗原的S1蛋白基因-T2A-N蛋白基因。
  4. 如权利要求3所述的用于预防新型冠状病毒的的生物制品，其特征在于，所述S1蛋白基因和/或N蛋白基因为人工合成且经密码子优化后的新型冠状病毒的基因序列。
  5. 如权利要求4所述的用于预防新型冠状病毒的的生物制品，其特征在于，所述S1蛋白基因的核苷酸序列如SEQ ID NO.1所示。
  6. 如权利要求4所述的用于预防新型冠状病毒的的生物制品，其特征在于，所述N蛋白基因的核苷酸序列如SEQ ID NO.2所示。
  7. 如权利要求3所述的用于预防新型冠状病毒的的生物制品，其特征在于，所述抗原蛋白基因表达盒的核苷酸序列如SEQ ID NO.3所示。
  8. 如权利要求3所述的用于预防新型冠状病毒的的生物制品，其特征在于，所述抗原蛋白基因表达盒的核苷酸序列如SEQ ID NO.4所示。
  9. 如权利要求1所述的用于预防新型冠状病毒的的生物制品，其特征在于，所述表达载体为E1和E3基因缺失的人5型腺病毒或质粒。
  10. 如权利要求1至9任一项所述的用于预防新型冠状病毒的生物制品，其特征在于，该生物制品为基因疫苗或基因药物。
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