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Severe acute respiratory syndrome coronavirus 2 vaccine based on novel adenovirus vectors Sad23L and/or Ad49L

An adenovirus, recombinant adenovirus technology, applied in the direction of viruses/phages, microorganism-based methods, viruses, etc., can solve problems such as limiting the application of adenovirus vectors

Active Publication Date: 2020-10-16
广州佰芮慷生物科技有限公司
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, the pre-existing high level of immune response against common human adenoviruses (Ad5 or Ad2) in the population limits the application of adenoviral vectors.

Method used

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  • Severe acute respiratory syndrome coronavirus 2 vaccine based on novel adenovirus vectors Sad23L and/or Ad49L
  • Severe acute respiratory syndrome coronavirus 2 vaccine based on novel adenovirus vectors Sad23L and/or Ad49L
  • Severe acute respiratory syndrome coronavirus 2 vaccine based on novel adenovirus vectors Sad23L and/or Ad49L

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0050] Example 1 Preparation of novel coronavirus vaccine based on replication-deficient adenovirus vectors Sad23L and Ad49L

[0051] 1. Codon optimization and acquisition of foreign gene S.

[0052] The gene sequence of the spike glycoprotein (S) of SARS-CoV-2 is derived from the new coronavirus strain (GenBank no.MN908947.3), and the codons are optimized using the software Upgene, making the foreign gene more suitable for use in mammals The cells were expressed, and the optimized gene sequence of S was shown in SEQ ID NO: 1, which was synthesized at Huada Gene to obtain the plasmid pMV-nCoV-S with the optimized foreign gene sequence.

[0053] 2. Construction of recombinant adenovirus vector and packaging of virus.

[0054] 2.1 Construction of the shuttle plasmid pShuttle2-CMV-S

[0055] The plasmid pMV-nCoV-S containing the gene sequence S synthesized by the whole gene was used EcoRI and BamHI Carry out double enzyme digestion, recover the digestion product, connect th...

Embodiment 2

[0073] Example 2 Immunological evaluation of novel coronavirus pneumonia COVID-19 vaccine Sad23L-nCoV-S on mouse model

[0074] 1. Evaluation of specific humoral immunity induced by recombinant adenovirus vaccine Sad23L-nCoV-S

[0075] 1.1 Inoculation titer and site of vaccine immunization

[0076] SPF female 5-week-old C57BL / 6 mice were purchased from the Animal Center of Southern Medical University and bred in the Animal Center of Southern Hospital. All animal husbandry and experiments complied with national and institutional animal welfare regulations. The injection volume was 100 μl per mouse. After four weeks of immunization, eyeball blood was collected, and the serum was separated to determine the levels of binding antibodies and neutralizing antibodies. The grouping and immunization of mice are shown in Table 1:

[0077] Table 1: Grouping of Sad23L-nCoV-S immunized mice, immunization dose and inoculation site

[0078]

[0079] 1.2 Levels of binding antibodies spe...

Embodiment 3

[0100] Example 3 Immunological Evaluation of New Coronavirus Pneumonia COVID-19 Vaccine Ad49L-nCoV-S on Mouse Model

[0101] 1. Evaluation of specific humoral immunity induced by recombinant adenovirus vaccine Ad49L-nCoV-S

[0102] 1.1 Inoculation titer and site of vaccine immunization

[0103] SPF female 5-week-old C57BL / 6 mice were purchased from the Animal Center of Southern Medical University and bred in the Animal Center of Southern Hospital. All animal husbandry and experiments complied with national and institutional animal welfare regulations. The injection volume was 100 μl per mouse. After four weeks of immunization, eyeball blood was collected, and the serum was separated to determine the levels of binding antibodies and neutralizing antibodies. The grouping and immunization of mice are shown in Table 2:

[0104] Table 2: Ad49L-nCoV-S immunized mice grouping, immunization dose and inoculation site

[0105]

[0106] 1.2 Levels of binding antibodies specific to...

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Abstract

The invention discloses a corona virus disease COVID-19 vaccine based on novel adenovirus vectors Sad23L and / or Ad49L. An exogenous gene is optimized to enable the expression of a foreign protein to be obviously increased, and besides, human rare or monkey adenoviruses are further attempted to be used as vectors, and a prestored immune reaction for common adenoviruses is avoided. The obtained corona virus disease COVID-19 vaccine can be induced in an animal body to generate high-level body fluid and cellular immunity, and after animal immunization, side effects do not occur. The corona virus disease COVID-19 vaccine is safe and effective, and can be quickly and massively prepared, and therefore, the corona virus disease COVID-19 vaccine is a candidata vaccine for preventing SARS-CoV-2.

Description

technical field [0001] The invention belongs to the field of biotechnology, and in particular relates to a novel coronavirus pneumonia COVID-19 vaccine developed based on novel adenovirus vectors Sad23L and / or Ad49L. Background technique [0002] The causative agent of coronavirus disease 2019 (COVID-19) was identified as novel coronavirus (severe acute respiratory syndrome coronavirus 2, SARS-CoV-2), which is similar to severe acute respiratory syndrome coronavirus (SARS-CoV, about 79%) and Middle East Respiratory Syndrome Coronavirus (MERA-CoV, about 50%) of Betacoronavirus (Betacoronavirus), Sarbecovirus subgenus of the novel coronavirus. The person-to-person transmission ability of SARS-CoV-2 is stronger than that of SARS-CoV and MERS-CoV, but its pathogenicity is lower than that of SARS-CoV, and there are people with mild or no obvious clinical symptoms. In view of the existence of occult carriers, it can be speculated that in the state of first-level response measures...

Claims

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Application Information

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IPC IPC(8): C12N15/50C07K14/165C12N15/861C12N7/01A61K39/215A61P31/14C12R1/93
CPCA61K39/12A61P31/14C07K14/005C12N7/00C12N15/86C12N2710/10321C12N2710/10343C12N2710/10351C12N2770/20022C12N2770/20034
Inventor 罗升学刘博超张攀丽
Owner 广州佰芮慷生物科技有限公司
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