Six broad-spectrum neutralizing antibodies against SARS-CoV-2 and their applications

Abstract

This invention discloses six broad-spectrum neutralizing antibodies against SARS-CoV-2 and their applications. This invention provides six IgG antibodies: HX001-020, HX001-013, HX001-015, HX001-024, HX001-033, and HX001-034. This invention protects any one or any combination of the six IgG antibodies. The CDR1, CDR2, and CDR3 in the heavy chain variable region of HX001-020, HX001-013, HX001-015, HX001-024, HX001-033, and HX001-034 are, respectively, positions 45-52, 70-77, and 116-130 of sequences 7, 3, 5, 9, 11, and 13. In the variable region of the light chain, CDR1, CDR2, and CDR3 are located at positions 46-51, 69-71, and 108-116 from the N-terminus of sequence 1, respectively. The six compounds provided by this invention have a broad-spectrum neutralizing effect against SARS-CoV-2.

Description

Technical Field

[0001] This invention belongs to the field of biotechnology and relates to six broad-spectrum neutralizing antibodies against SARS-CoV-2 and their applications. Background Technology

[0002] The novel coronavirus (SARS-CoV-2) belongs to the β-coronavirus genus. COVID-19 is caused by infection with the novel coronavirus. The main symptoms are low-grade fever, fatigue, and dry cough. A few patients may also experience upper respiratory and digestive symptoms such as nasal congestion, runny nose, and diarrhea. Severe cases may develop into acute respiratory distress syndrome, septic shock, metabolic acidosis, coagulation dysfunction, and multiple organ failure.

[0003] During its transmission, SARS-CoV-2 has undergone numerous mutations, enhancing its transmissibility and causing immune evasion. Currently, the main circulating naturally occurring mutant strains include the Alpha strain found in the UK, the Beta strain found in South Africa, the Gamma strain found in Brazil, and the Delta strain found in India. The emergence of these mutant strains has impacted vaccine immunization against the novel coronavirus. Inactivated vaccine serum showed a 50% reduction in neutralizing activity against the South African mutant strain, while mRNA vaccine serum showed approximately a 10-fold reduction in neutralizing activity. Many monoclonal antibodies have also shown weakened or completely lost their neutralizing activity against the mutant strains.

[0004] Monoclonal antibodies can be mass-produced industrially. Their high affinity and specificity for antigen binding significantly reduce adverse reactions in clinical applications. Furthermore, antibody molecules can be modified to increase their antiviral efficacy. Antibodies, with their specificity and flexibility of use, are a very promising tool in the treatment of infectious diseases. Currently, there is an urgent need to address the challenges posed by naturally occurring mutant strains of the novel coronavirus. Developing broad-spectrum human monoclonal antibodies against the novel coronavirus will provide more effective prevention and treatment for infections caused by these naturally occurring mutant strains. Summary of the Invention

[0005] The purpose of this invention is to provide six broad-spectrum neutralizing antibodies against SARS-CoV-2 and their applications.

[0006] This invention provides six IgG antibodies. The six IgG antibodies are: HX001-020 antibody, HX001-013 antibody, HX001-015 antibody, HX001-024 antibody, HX001-033 antibody, and HX001-034 antibody.

[0007] This invention protects any one of the six IgG antibodies.

[0008] This invention also protects any combination of the six IgG antibodies.

[0009] Specifically, this invention protects the combination of HX001-020 antibody with any one, any two, any three, or any four of the remaining five antibodies.

[0010] Specifically, this invention protects combinations of the six antibodies.

[0011] The HX001-020 antibody is composed of a light chain and a heavy chain; the CDR1, CDR2, and CDR3 in the variable region of the heavy chain are amino acid residues from the N-terminus of sequence 7 in the sequence listing, namely positions 45-52, 70-77, and 116-130, respectively; the CDR1, CDR2, and CDR3 in the variable region of the light chain are amino acid residues from the N-terminus of sequence 1 in the sequence listing, namely positions 46-51, 69-71, and 108-116, respectively.

[0012] The HX001-013 antibody is composed of a light chain and a heavy chain; the CDR1, CDR2, and CDR3 in the variable region of the heavy chain are amino acid residues from the N-terminus of sequence 3 in the sequence listing, namely positions 45-52, 70-77, and 116-130 respectively; the CDR1, CDR2, and CDR3 in the variable region of the light chain are amino acid residues from the N-terminus of sequence 1 in the sequence listing, namely positions 46-51, 69-71, and 108-116 respectively.

[0013] The HX001-015 antibody is composed of a light chain and a heavy chain; the CDR1, CDR2, and CDR3 in the variable region of the heavy chain are amino acid residues from the N-terminus of sequence 5 in the sequence listing, namely positions 45-52, 70-77, and 116-130, respectively; the CDR1, CDR2, and CDR3 in the variable region of the light chain are amino acid residues from the N-terminus of sequence 1 in the sequence listing, namely positions 46-51, 69-71, and 108-116, respectively.

[0014] The HX001-024 antibody is composed of a light chain and a heavy chain; the CDR1, CDR2, and CDR3 in the variable region of the heavy chain are amino acid residues from the N-terminus of sequence 9 in the sequence listing, namely positions 45-52, 70-77, and 116-130, respectively; the CDR1, CDR2, and CDR3 in the variable region of the light chain are amino acid residues from the N-terminus of sequence 1 in the sequence listing, namely positions 46-51, 69-71, and 108-116, respectively.

[0015] The HX001-033 antibody is composed of a light chain and a heavy chain; the CDR1, CDR2, and CDR3 in the variable region of the heavy chain are amino acid residues from the N-terminus of sequence 11 in the sequence listing, namely positions 45-52, 70-77, and 116-130 respectively; the CDR1, CDR2, and CDR3 in the variable region of the light chain are amino acid residues from the N-terminus of sequence 1 in the sequence listing, namely positions 46-51, 69-71, and 108-116 respectively.

[0016] The HX001-034 antibody is composed of a light chain and a heavy chain; the CDR1, CDR2, and CDR3 in the variable region of the heavy chain are amino acid residues from the N-terminus of sequence 13 in the sequence listing, namely positions 45-52, 70-77, and 116-130 respectively; the CDR1, CDR2, and CDR3 in the variable region of the light chain are amino acid residues from the N-terminus of sequence 1 in the sequence listing, namely positions 46-51, 69-71, and 108-116 respectively.

[0017] The HX001-020 antibody has a heavy chain variable region consisting of amino acid residues from the N-terminus of sequence 7 in the sequence listing, from position 20 to 142; and a light chain variable region consisting of amino acid residues from the N-terminus of sequence 1 in the sequence listing, from position 20 to 126.

[0018] The HX001-013 antibody has a heavy chain variable region consisting of amino acid residues from the N-terminus of sequence 3 in the sequence listing, from position 20 to 142; and a light chain variable region consisting of amino acid residues from the N-terminus of sequence 1 in the sequence listing, from position 20 to 126.

[0019] The HX001-015 antibody has a heavy chain variable region consisting of amino acid residues from the N-terminus of sequence 5 in the sequence listing, from position 20 to 142; and a light chain variable region consisting of amino acid residues from the N-terminus of sequence 1 in the sequence listing, from position 20 to 126.

[0020] The HX001-024 antibody has a heavy chain variable region consisting of amino acid residues from the N-terminus of sequence 9 in the sequence listing, from position 20 to 142; and a light chain variable region consisting of amino acid residues from the N-terminus of sequence 1 in the sequence listing, from position 20 to 126.

[0021] The HX001-033 antibody has a heavy chain variable region consisting of amino acid residues from the N-terminus of sequence 11 in the sequence listing, from position 20 to 142; and a light chain variable region consisting of amino acid residues from the N-terminus of sequence 1 in the sequence listing, from position 20 to 126.

[0022] The HX001-034 antibody has a heavy chain variable region consisting of amino acid residues from the N-terminus of sequence 13 in the sequence listing, from position 20 to 142; and a light chain variable region consisting of amino acid residues from the N-terminus of sequence 1 in the sequence listing, from position 20 to 126.

[0023] The HX001-020 antibody has its heavy chain as shown in sequence 7 of the sequence listing and its light chain as shown in sequence 1 of the sequence listing.

[0024] The HX001-013 antibody has its heavy chain as shown in sequence 3 of the sequence listing and its light chain as shown in sequence 1 of the sequence listing.

[0025] The HX001-015 antibody has its heavy chain as shown in sequence 5 of the sequence listing; and its light chain as shown in sequence 1 of the sequence listing.

[0026] The HX001-024 antibody has its heavy chain as shown in sequence 9 of the sequence listing; and its light chain as shown in sequence 1 of the sequence listing.

[0027] The HX001-033 antibody has its heavy chain as shown in sequence 11 of the sequence listing; and its light chain as shown in sequence 1 of the sequence listing.

[0028] The HX001-034 antibody has its heavy chain as shown in sequence 13 of the sequence listing; and its light chain as shown in sequence 1 of the sequence listing.

[0029] This invention also protects any one or any combination of the six genes;

[0030] The six genes encode six IgG antibodies;

[0031] The six IgG antibodies are: HX001-020 antibody, HX001-013 antibody, HX001-015 antibody, HX001-024 antibody, HX001-033 antibody and HX001-034 antibody.

[0032] The gene encoding the HX001-020 antibody consists of a gene encoding the heavy chain and a gene encoding the light chain; the gene encoding the heavy chain is shown as nucleotides 892-2307 in sequence 8 of the sequence listing; the gene encoding the light chain is shown as nucleotides 1038-1739 in sequence 2 of the sequence listing.

[0033] The gene encoding the HX001-013 antibody consists of a gene encoding the heavy chain and a gene encoding the light chain; the gene encoding the heavy chain is shown as nucleotides 892-2307 in sequence 4 of the sequence listing; the gene encoding the light chain is shown as nucleotides 1038-1739 in sequence 2 of the sequence listing.

[0034] The gene encoding the HX001-015 antibody consists of a gene encoding the heavy chain and a gene encoding the light chain; the gene encoding the heavy chain is shown as nucleotides 892-2307 in sequence 6 of the sequence listing; the gene encoding the light chain is shown as nucleotides 1038-1739 in sequence 2 of the sequence listing.

[0035] The gene encoding the HX001-024 antibody consists of a gene encoding the heavy chain and a gene encoding the light chain; the gene encoding the heavy chain is shown as nucleotides 892-2307 in sequence 10 of the sequence listing; the gene encoding the light chain is shown as nucleotides 1038-1739 in sequence 2 of the sequence listing.

[0036] The gene encoding the HX001-033 antibody consists of a gene encoding the heavy chain and a gene encoding the light chain; the gene encoding the heavy chain is shown as nucleotides 892-2307 in sequence 12 of the sequence listing; the gene encoding the light chain is shown as nucleotides 1038-1739 in sequence 2 of the sequence listing.

[0037] The gene encoding the HX001-034 antibody consists of a gene encoding the heavy chain and a gene encoding the light chain; the gene encoding the heavy chain is shown as nucleotides 892-2307 in sequence 14 of the sequence listing; the gene encoding the light chain is shown as nucleotides 1038-1739 in sequence 2 of the sequence listing.

[0038] The present invention also protects the use of any of the above-described IgG antibodies or combinations of IgG antibodies in the preparation of medicaments for inhibiting the novel coronavirus.

[0039] The present invention also protects a drug for inhibiting the novel coronavirus, wherein the active ingredient is any of the IgG antibodies or combinations of IgG antibodies described above.

[0040] The present invention also protects the use of any of the above-described IgG antibodies or combinations of IgG antibodies in the preparation of medicaments for neutralizing the novel coronavirus.

[0041] The present invention also protects a drug for neutralizing the novel coronavirus, the active ingredient of which is any of the IgG antibodies or combinations of IgG antibodies described above.

[0042] The present invention also protects the use of any of the above-described IgG antibodies or combinations of IgG antibodies in the preparation of medicaments for the prevention and / or treatment of COVID-19.

[0043] The present invention also protects a medicament for the prevention and / or treatment of COVID-19, wherein the active ingredient is any of the IgG antibodies or combinations of IgG antibodies described above.

[0044] The COVID-19 pandemic described above is caused by the novel coronavirus.

[0045] The novel coronavirus mentioned above is either a wild-type novel coronavirus or a naturally occurring variant of the wild-type novel coronavirus.

[0046] The novel coronavirus mentioned above is a wild-type novel coronavirus, a novel coronavirus Alpha strain, a novel coronavirus Beta strain, a novel coronavirus Gamma strain, a novel coronavirus Delta strain, a novel coronavirus DeltaPlus strain, a novel coronavirus Kappa strain, a novel coronavirus Epsilon strain, a novel coronavirus Eta strain, a novel coronavirus Iota strain, or a novel coronavirus N439K strain.

[0047] In preliminary experiments, the inventors of this invention used the spike protein of the novel coronavirus as bait, screening antibody-generating memory B cells from peripheral blood mononuclear cells of infected individuals to obtain monoclonal antibodies that specifically bind to the spike protein, named antibody P36-5D2. Based on antibody P36-5D2, the inventors further obtained six optimized antibodies through extensive preliminary optimization experiments. The six antibodies provided by this invention have broad-spectrum neutralizing effects against SARS-CoV-2, exhibiting strong neutralizing capabilities against both wild-type novel coronavirus and naturally occurring variants. This invention has significant application value for the prevention and control of the novel coronavirus and will have profound social implications. Attached Figure Description

[0048] Figure 1 This is a schematic diagram of the amino acid sequence elements of the six heavy chains.

[0049] Figure 2 This is a schematic diagram of the differential regions between the amino acid sequences of the six heavy chains.

[0050] Figure 3 This is the result of Example 2. Detailed Implementation

[0051] The present invention will now be described in further detail with reference to specific embodiments. The given embodiments are merely illustrative of the invention and not intended to limit its scope. The embodiments provided below can serve as a guide for further improvements by those skilled in the art and do not constitute a limitation on the invention in any way.

[0052] Unless otherwise specified, the experimental methods used in the following examples are conventional methods, performed according to the techniques or conditions described in the literature in this field or according to the product instructions. Unless otherwise specified, the materials and reagents used in the following examples are commercially available. hACE2-hela cells (ie "HeLa cell lines stably expressing the ACE2" in the literature molecules"), recorded in the following literature: Wang, R., Zhang, Q., Ge, J., Ren, W., Zhang, R., Lan, J., Ju, B., Su, B., Yu, F., Chen, P .,Liao,H.,Feng,Y.,Li,X.,Shi,X.,Zhang,Z.,Zhang,F.,Ding,Q.,Zhang,T.,Wang,X.&Zhang,L.Analysis of SARS-CoV-2 variantmutations reveals neutralization escape mechanisms and the ability to use ACE2 receptors from additional species. Immunity 54, 1611-1621.e1615, doi:10.1016 / j.immuni.2021.06.003 (2021). Plasmid pcDNA3.1(+): Invitrogen, catalog number V790-20. 293T cells: Gader, CRL-11268.

[0053] Unless otherwise specified, the quantitative experiments in the following examples are all repeated three times, and the results are averaged.

[0054] Experimental Example 1: Preparation of Antibodies

[0055] I. Preparation of recombinant plasmids

[0056] The DNA molecule shown in Sequence 2 of the sequence listing was inserted into the pLB-simple vector to obtain the recombinant plasmid. This recombinant plasmid has been verified by sequencing. This recombinant plasmid is also named the light chain expression plasmid. In Sequence 2 of the sequence listing, nucleotides 1-1037 form the promoter, nucleotides 1038-1739 form the light chain coding region, and nucleotides 1740-1887 form the terminator. The DNA molecule shown in Sequence 2 of the sequence listing expresses the light chain shown in Sequence 1 of the sequence listing. In Sequence 1 of the sequence listing, amino acid residues 20-126 are the variable region (CDR1, CDR2, and CDR3 are shown as positions 46-51, 69-71, and 108-116 of Sequence 1 of the sequence listing, respectively), and amino acid residues 127-233 are the constant region.

[0057] The DNA molecule shown in Sequence 4 of the sequence listing was inserted into the pLB-simple vector to obtain recombinant plasmid-013. This recombinant plasmid has been verified by sequencing. This recombinant plasmid is also named heavy chain expression plasmid-013. In Sequence 4 of the sequence listing, nucleotides 1-891 are the promoter, nucleotides 892-2307 are the heavy chain coding region, and nucleotides 2308-2453 are the terminator. The DNA molecule shown in Sequence 4 of the sequence listing expresses the heavy chain-013 shown in Sequence 3 of the sequence listing. In Sequence 3 of the sequence listing, amino acid residues 20-142 are the variable region (CDR1, CDR2, and CDR3 are shown as positions 45-52, 70-77, and 116-130 of Sequence 3 of the sequence listing, respectively), and amino acid residues 143-471 are the constant region.

[0058] The DNA molecule shown in Sequence 6 of the sequence listing was inserted into the pLB-simple vector to obtain recombinant plasmid-015. This recombinant plasmid has been verified by sequencing. This recombinant plasmid is also named heavy chain expression plasmid-015. In Sequence 6 of the sequence listing, nucleotides 1-891 are the promoter, nucleotides 892-2307 are the heavy chain coding region, and nucleotides 2308-2453 are the terminator. The DNA molecule shown in Sequence 6 of the sequence listing expresses the heavy chain-015 shown in Sequence 5 of the sequence listing. In Sequence 5 of the sequence listing, amino acid residues 20-142 are the variable region (CDR1, CDR2, and CDR3 are shown as positions 45-52, 70-77, and 116-130 of Sequence 5 of the sequence listing, respectively), and amino acid residues 143-471 are the constant region.

[0059] The DNA molecule shown in Sequence 8 of the sequence listing was inserted into the pLB-simple vector to obtain recombinant plasmid-020. This recombinant plasmid has been verified by sequencing. This recombinant plasmid is also named heavy chain expression plasmid-020. In Sequence 8 of the sequence listing, nucleotides 1-891 are the promoter, nucleotides 892-2307 are the heavy chain coding region, and nucleotides 2308-2453 are the terminator. The DNA molecule shown in Sequence 8 of the sequence listing expresses heavy chain-020 shown in Sequence 7 of the sequence listing. In Sequence 7 of the sequence listing, amino acid residues 20-142 are the variable region (CDR1, CDR2, and CDR3 are shown as positions 45-52, 70-77, and 116-130 of Sequence 7 of the sequence listing, respectively), and amino acid residues 143-471 are the constant region.

[0060] The DNA molecule shown in Sequence 10 of the sequence listing was inserted into the pLB-simple Vector to obtain recombinant plasmid-024. This recombinant plasmid has been verified by sequencing. This recombinant plasmid is also named heavy chain expression plasmid-024. In Sequence 10 of the sequence listing, nucleotides 1-891 are the promoter, nucleotides 892-2307 are the heavy chain coding region, and nucleotides 2308-2453 are the terminator. The DNA molecule shown in Sequence 10 of the sequence listing expresses heavy chain-024 shown in Sequence 9 of the sequence listing. In Sequence 9 of the sequence listing, amino acid residues 20-142 are the variable region (CDR1, CDR2, and CDR3 are shown as positions 45-52, 70-77, and 116-130 of Sequence 9 of the sequence listing, respectively), and amino acid residues 143-471 are the constant region.

[0061] The DNA molecule shown in Sequence 12 of the sequence listing was inserted into the pLB-simple vector to obtain recombinant plasmid-033. This recombinant plasmid has been verified by sequencing. This recombinant plasmid is also named heavy chain expression plasmid-033. In Sequence 12 of the sequence listing, nucleotides 1-891 are the promoter, nucleotides 892-2307 are the heavy chain coding region, and nucleotides 2308-2453 are the terminator. The DNA molecule shown in Sequence 12 of the sequence listing expresses heavy chain-033 shown in Sequence 11 of the sequence listing. In Sequence 11 of the sequence listing, amino acid residues 20-142 are the variable region (CDR1, CDR2, and CDR3 are shown as positions 45-52, 70-77, and 116-130 of Sequence 11 of the sequence listing, respectively), and amino acid residues 143-471 are the constant region.

[0062] The DNA molecule shown in Sequence 14 of the sequence listing was inserted into the pLB-simple Vector to obtain recombinant plasmid-034. This recombinant plasmid has been verified by sequencing. This recombinant plasmid is also named heavy chain expression plasmid-034. In Sequence 14 of the sequence listing, nucleotides 1-891 are the promoter, nucleotides 892-2307 are the heavy chain coding region, and nucleotides 2308-2453 are the terminator. The DNA molecule shown in Sequence 14 of the sequence listing expresses the heavy chain-034 shown in Sequence 13 of the sequence listing. In Sequence 13 of the sequence listing, amino acid residues 20-142 are the variable region (CDR1, CDR2, and CDR3 are shown as positions 45-52, 70-77, and 116-130 of Sequence 13 of the sequence listing, respectively), and amino acid residues 143-471 are the constant region.

[0063] The DNA molecule shown in Sequence 16 of the sequence listing was inserted into the pLB-simple vector to obtain the recombinant plasmid P36-5D2. This recombinant plasmid has been verified by sequencing. This recombinant plasmid is also named heavy chain expression plasmid P36-5D2. In Sequence 16 of the sequence listing, nucleotides 1-891 are the promoter, nucleotides 892-2307 are the heavy chain coding region, and nucleotides 2308-2453 are the terminator. The DNA molecule shown in Sequence 16 of the sequence listing expresses the heavy chain P36-5D2 shown in Sequence 15 of the sequence listing. In Sequence 15 of the sequence listing, amino acid residues 20-142 are the variable region (CDR1, CDR2, and CDR3 are shown as positions 45-52, 70-77, and 116-130 of Sequence 15 of the sequence listing, respectively), and amino acid residues 143-471 are the constant region.

[0064] A schematic diagram of the amino acid sequence elements of the 6 heavy chains is shown below. Figure 1 . Figure 1 In the text, the underlined areas are variable areas, and the yellow background indicates CorelDRAW (CorelDRAW) text.

[0065] A schematic diagram of the differential regions between the amino acid sequences of the six heavy chains is shown below. Figure 2 .

[0066] pLB-simple Vector is a component of Tiangen Biotech's pLB zero-background rapid ligation kit (catalog number VT206), available at http: / / www.tiangen.com / ?productShow / t1 / 6 / id / 308.html.

[0067] II. Preparation of HX001-013 antibody

[0068] 1. The light chain expression plasmid prepared in step 1 and the heavy chain expression plasmid-013 prepared in step 1 were co-transfected into 293T cells, and then cultured in DMEM medium containing 2% fetal bovine serum for 72 h. After centrifugation at 4℃ and 4000 rpm for 30 min, the supernatant was collected.

[0069] 2. Affinity chromatography

[0070] Affinity chromatography column specifications: length 3cm, inner diameter 1cm;

[0071] Affinity chromatography column packing material: protein A beads (Thermo, catalog number 10006D);

[0072] Perform the following steps in sequence: ① Load 300 mL of the supernatant obtained in step 1 onto an affinity chromatography column and incubate at 4 °C for 16 hours; ② Wash the column with 60 mL of binding buffer; ③ Elute the target protein with 30 mL of elution buffer and collect the post-column solution.

[0073] Binding buffer: Dissolve 112.6g of glycine and 175.2g of sodium chloride in water and bring the volume to 1L. Adjust the pH to 8.0 with sodium hydroxide.

[0074] Elution buffer: Dissolve 7.5g of glycine in water and bring the volume to 500mL. Adjust the pH to 3.0 with hydrochloric acid.

[0075] 3. Take the post-column solution obtained in step 2, concentrate it with an ultrafiltration concentrator and replace the system with PBS buffer (pH 7.2, 10mM) to obtain 1mL of antibody solution (antibody concentration is about 2mg / mL), which is called HX001-013 antibody solution.

[0076] III. Preparation of HX001-015 antibody

[0077] Replace heavy chain expression plasmid-013 with heavy chain expression plasmid-015, and follow the same procedure as in step two to obtain the HX001-015 antibody solution.

[0078] IV. Preparation of HX001-020 antibody

[0079] Replace heavy chain expression plasmid-013 with heavy chain expression plasmid-020, and follow the same procedure as in step two to obtain the HX001-020 antibody solution.

[0080] V. Preparation of HX001-024 antibody

[0081] Replace heavy chain expression plasmid-013 with heavy chain expression plasmid-024, and follow the same procedure as in step two to obtain the HX001-024 antibody solution.

[0082] VI. Preparation of HX001-033 antibody

[0083] Replace heavy chain expression plasmid-013 with heavy chain expression plasmid-033, and follow the same procedure as in step two to obtain HX001-033 antibody solution.

[0084] VII. Preparation of HX001-034 antibody

[0085] Replace heavy chain expression plasmid-013 with heavy chain expression plasmid-034, and follow the same procedure as in step two to obtain HX001-034 antibody solution.

[0086] 8. Preparation of P36-5D2 antibody

[0087] Replace the heavy chain expression plasmid-013 with the heavy chain expression plasmid P36-5D2, and follow the same procedure as in step two to obtain the P36-5D2 antibody solution.

[0088] Example 2: Detection of antibody affinity for novel coronavirus RBD

[0089] Detection Principle: Surface plasmon resonance (SPR) is an optical physical sensing technology capable of analyzing the concentration, affinity, kinetic constants, and specificity of various small organic molecules, proteins, nucleic acids, and carbohydrates. First, a layer of molecules is immobilized on the chip surface. Then, the sample to be tested flows through the chip surface. Molecules in the sample that can interact with the molecules on the chip surface cause changes in the surface refractive index, ultimately leading to a change in the surface plasmon resonance angle, which is then detected.

[0090] The test antibodies were: HX001-013 antibody, HX001-015 antibody, HX001-020 antibody, HX001-024 antibody, HX001-033 antibody, HX001-034 antibody, or P36-5D2 antibody. All antibodies were provided by the corresponding antibody solutions prepared in Example 1.

[0091] The binding kinetics of the test antibody to SARS-CoV-2 RBD were detected using a Biocore 8K instrument. A multi-cycle kinetics using capture program was run. Recombinant protein A was covalently immobilized onto the CM5 sensor chip via amine groups in 10 mM sodium acetate buffer (pH 4.5). The final response units (RU) at a capture antibody concentration of 5 μg were approximately 700. The test antibody was captured on the sensor chip coated with recombinant protein A, and then serially diluted SARS-CoV-2 RBD monomers (as shown in Sequence 17 of the sequence listing) were flowed through the sensor chip system. The SARS-CoV-2 RBD concentration started at 200 nM, with two-fold serial dilutions, eight dilutions in total, and a 0 concentration control. Three replicates were set for each concentration and the 0 concentration control. The antigen binding time was generally 120 s, and the dissociation time was adjusted accordingly: 250 s for weak antigen-antibody binding and 1000 s for strong antigen-antibody binding. Running buffer HBS-EP: contains 0.15M NaCl and 0.05% (v / v) Tween-20, with the balance being 0.01M HEPES buffer at pH 7.4. Regeneration buffer (pH 1.5): contains 7.507g glycine per 500ml, with the balance being water.

[0092] Data analysis of affinity was performed using multi-cycle kinetics with the capture-Evaluation method. After confirming that there were no issues with QC, model fitting (1:1 binding) was performed to calculate the binding dissociation constant (KD) value.

[0093] See results Figure 3 . Figure 3 In the graph, the vertical axis represents the response unit (RU), and the horizontal axis represents time (seconds). Compared to the P36-5D2 antibody, the six optimized antibodies (HX001-013, HX001-015, HX001-020, HX001-024, HX001-033, and HX001-034) showed a 20-50 fold increase in affinity for SARS-CoV-2 RBD.

[0094] Example 3: Detection of neutralizing activity of antibodies against novel coronavirus pseudovirus

[0095] The test antibodies were: HX001-013 antibody, HX001-015 antibody, HX001-020 antibody, HX001-024 antibody, HX001-033 antibody, HX001-034 antibody, or P36-5D2 antibody, or REGN10987 antibody. HX001-013, HX001-015, HX001-020, HX001-024, HX001-033, HX001-034, and P36-5D2 antibodies were all provided by the corresponding antibody solutions prepared in Example 1. REGN10987 antibody is a neutralizing antibody against SARS-CoV-2 (novel coronavirus) and has received emergency use authorization from the US FDA. The REGN10987 antibody was prepared according to the method in Example 1. The only difference between the REGN10987 antibody and the P36-5D2 antibody is the heavy chain variable region and the light chain variable region. The heavy chain variable region of the REGN10987 antibody is shown in Sequence 20 of the sequence listing, and the light chain variable region of the REGN10987 antibody is shown in Sequence 21 of the sequence listing.

[0096] The membrane proteins of 10 novel coronavirus strains originate from the following sources:

[0097] Wild-type novel coronavirus (Genbank: MN908947.3);

[0098] The novel coronavirus Alpha strain (GISAID: EPI_ISL_601443) contains 9 mutations: 69-70del, 144del, N501Y, A570D, D614G, P681H, T716I, S982A, and D1118H; B.1.1.7;

[0099] The novel coronavirus Beta strain (GISAID: EPI_ISL_700450) contains 10 mutations: L18F, D80A, D215G, 242-244del, S305T, K417N, E484K, N510Y, D614G, A701V; B.1.351;

[0100] The novel coronavirus Gamma strain (GISAID: EPI_ISL_792681) contains 12 mutations: L18F, T20N, P26S, D138Y, R190S, K417T, E484K, N501Y, D614G, H655Y, T1027I, V1176F; P.1.

[0101] The novel coronavirus Delta strain (GISAID: EPI_ISL_1534938) contains 10 mutations: T19R, G142D, 156-157del, R158G, A222V, L452R, T478K, D614G, P681R, and D950N; B.1.617.2;

[0102] The novel coronavirus Delta Plus strain (GISAID: EPI_ISL_3019629) contains 11 mutations: T19R, G142D, 156-157del, R158G, A222V, K417N, L452R, T478K, D614G, P681R, and D950N; B.1.617.2.1;

[0103] The novel coronavirus Kappa strain (GISAID: EPI_ISL_1384866) contains 8 mutations: T95I, G142D, E154L, L452R, E484Q, D614G, P681R, N1071H; B.1.617.1;

[0104] The novel coronavirus Epsilon strain (GISAID: EPI_ISL_2922315) contains four mutations: S13I, W152C, L452R, D614G; B.1.429;

[0105] The novel coronavirus Eta strain (GISAID: EPI_ISL_2885901) contains 8 mutations: Q52R, A67V, 69-70del, 144del, E484K, D614G, Q677H, F888L; B.1.525;

[0106] The novel coronavirus Iota strain (GISAID: EPI_ISL_2922249) contains 6 mutations: L5F, T95I, D253G, E484K, D614G, A701V; B.1.526;

[0107] The novel coronavirus strain N439K contains one mutation: N439K.

[0108] I. Preparation of Novel Coronavirus Pseudovirus

[0109] Co-transfection of 293T cells with a plasmid expressing the SARS-CoV-2 membrane protein and a backbone plasmid pNL4-3R-E-luciferase yielded an infectious but non-replicating pseudovirus of SARS-CoV-2, with infectivity similar to that of live SARS-CoV-2. The backbone plasmid pNL4-3R-E-luciferase, i.e., the backbone plasmid pNL4-3R-E containing Luciferase (i.e., vector with the luciferase gene containing backbone pNL4-3R-E in the literature): Wang Q, Liu L, Ren W, Gettie A, Wang H, Liang Q, Shi X, Montefiori DC, Zhou T, Zhang L. Cell Rep. 2019.

[0110] The gene encoding the novel coronavirus membrane protein was inserted between the BamHII and EcoRI restriction sites of the pcDNA3.1(+) vector to obtain a plasmid expressing the novel coronavirus membrane protein. The plasmid expressing the novel coronavirus membrane protein and the backbone plasmid pNL4-3R-E-luciferase were co-transfected into 293T cells and incubated at 37°C (using DMEM medium containing 10% fetal bovine serum). The cell culture supernatant was collected 60 hours after transfection; this was the viral solution containing the novel coronavirus pseudovirus.

[0111] In the wild-type novel coronavirus, the membrane protein is shown in Sequence 18 of the sequence listing. When the gene encoding the novel coronavirus membrane protein is shown in Sequence 19 of the sequence listing (encoding the protein shown in Sequence 18 of the sequence listing), the above steps are performed to obtain a pseudovirus of the wild-type novel coronavirus. Based on sequence 19 in the sequence listing, various mutations were performed (the corresponding mutations of the novel coronavirus Alpha strain are shown in Table 1, the corresponding mutations of the novel coronavirus Beta strain are shown in Table 2, the corresponding mutations of the novel coronavirus Gamma strain are shown in Table 3, the corresponding mutations of the novel coronavirus Delta strain are shown in Table 4, the corresponding mutations of the novel coronavirus Delta Plus strain are shown in Table 5, the corresponding mutations of the novel coronavirus Kappa strain are shown in Table 6, the corresponding mutations of the novel coronavirus Epsilon strain are shown in Table 7, the corresponding mutations of the novel coronavirus Eta strain are shown in Table 8, the corresponding mutations of the novel coronavirus Iota strain are shown in Table 9, and the corresponding mutations of the novel coronavirus N439K strain are shown in Table 10). Then, as the coding gene of the novel coronavirus membrane protein, the above steps were performed to obtain novel coronavirus Alpha strain pseudovirus, novel coronavirus Beta strain pseudovirus, novel coronavirus Gamma strain pseudovirus, novel coronavirus Delta strain pseudovirus, novel coronavirus Delta Plus strain pseudovirus, novel coronavirus Kappa strain pseudovirus, novel coronavirus Epsilon strain pseudovirus, novel coronavirus Eta strain pseudovirus, novel coronavirus Iota strain pseudovirus, and novel coronavirus N439K strain pseudovirus.

[0112] Table 1

[0113] Protein mutation DNA mutation (corresponding to position 19 in sequence) 69-70del (missing two amino acid residues "HV") Missing bits 205-210 of "CACGTG" 144del (missing one amino acid residue "Y") The "TAT" character is missing in bits 430-432. N501Y (a mutation involving one amino acid residue) The "A" at position 1501 mutates to "T". A570D (a mutation involving one amino acid residue) The "C" at position 1709 mutates to "A". D614G (a single amino acid residue mutation) The "A" at position 1841 mutates to "G". P681H (a mutation involving one amino acid residue) The 2042nd position "C" mutates to "A". T716I (a single amino acid residue mutation) The "C" at position 2147 mutates to "T". S982A (a mutation involving one amino acid residue) The "AG" at positions 2944-2945 mutates to "GC". D1118H (a mutation involving one amino acid residue) The "G" at position 3352 mutates to "C".

[0114] Table 2

[0115] Protein mutation DNA mutation (corresponding to position 19 in sequence) L18F (a mutation involving one amino acid residue) The 52-bit "C" mutates to "T", and the 54-bit "G" mutates to "C". D80A (a mutation involving one amino acid residue) The "A" at position 239 mutates to "C". D215G (a mutation involving one amino acid residue) The 644-bit "A" mutates to "G". 242-244del (three amino acid residues missing) Missing bits 724-732 of "CTGGCCCTG" S305T (a mutation involving one amino acid residue) The 914th position "G" mutates to "C". K417N (a single amino acid residue mutation) The "G" at position 1251 mutates to "C". E484K (a single amino acid residue mutation) The "G" at position 1450 mutated to "A". N501Y (a mutation involving one amino acid residue) The "A" at position 1501 mutates to "T", and the "T" at position 1503 mutates to "C". D614G (a single amino acid residue mutation) The "A" at position 1841 mutates to "G". A701V (a mutation involving one amino acid residue) The "CC" at positions 2102-2103 mutates to "TG".

[0116] Table 3

[0117] Protein mutation DNA mutation (corresponding to position 19 in sequence) L18F (a mutation involving one amino acid residue) 52-bit C becomes T, 54-bit G becomes C. T20N (a mutation involving one amino acid residue) The "C" at position 59 mutates to "A". P26S (a mutation involving one amino acid residue) The "CCT" at positions 76-78 mutates to "AGC". D138Y (a mutation involving one amino acid residue) The 412-bit "G" mutates to "T". R190S (a single amino acid residue mutation) The "A" at position 570 mutates to "C". K417T (a single amino acid residue mutation) The "AG" at positions 1250-1251 mutates to "CC". E484K (a single amino acid residue mutation) The "G" at position 1450 mutated to "A". N501Y (a mutation involving one amino acid residue) The "A" at position 1501 mutates to "T". D614G (a single amino acid residue mutation) The "A" at position 1841 mutates to "G". H655Y (a mutation involving one amino acid residue) The "C" at position 1963 mutated to "T". T1027I (a single amino acid residue mutation) The 3080th position "C" mutates to "T". V1176F (a single amino acid residue mutation) The "G" at position 3526 mutates to "T", and the "G" at position 3528 mutates to "C".

[0118] Table 4

[0119] Protein mutation DNA mutation (corresponding to position 19 in sequence) T19R (a mutation involving one amino acid residue) The 55th position "A" mutates to "C", and the 56th position "C" mutates to "G". G142D (a single amino acid residue mutation) The 425th bit "G" mutates to "A". 156-157del (two amino acid residues missing) Missing bits 467-472 of "AGTTCC" R158G (a mutation involving one amino acid residue) The 474-bit "C" mutates to "T". A222V (a mutation involving one amino acid residue) The "C" at position 665 mutates to "T". L452R (a single amino acid residue mutation) The "T" at position 1355 mutates to "G". T478K (a single amino acid residue mutation) The "C" at position 1433 mutates to "A". D614G (a single amino acid residue mutation) The "A" at position 1841 mutates to "G". P681R (a mutation involving one amino acid residue) The "CCT" at positions 2041-2043 mutated to "AGA". D950N (a single amino acid residue mutation) The "G" at position 2848 mutates to "A", and the "C" at position 2850 mutates to "T".

[0120] Table 5

[0121] Protein mutation DNA mutation (corresponding to position 19 in sequence) T19R (a mutation involving one amino acid residue) The 55th position "A" mutates to "C", and the 56th position "C" mutates to "G". G142D (a single amino acid residue mutation) The 425th bit "G" mutates to "A". 156-157del (two amino acid residues missing) Missing bits 467-472 of "AGTTCC" R158G (a mutation involving one amino acid residue) The 474-bit "C" mutates to "T". A222V (a mutation involving one amino acid residue) The "C" at position 665 mutates to "T". K417N (a single amino acid residue mutation) The "AG" at positions 1250-1251 mutates to "CC". L452R (a single amino acid residue mutation) The "T" at position 1355 mutates to "G". T478K (a single amino acid residue mutation) The "C" at position 1433 mutates to "A". D614G (a single amino acid residue mutation) The "A" at position 1841 mutates to "G". P681R (a mutation involving one amino acid residue) The "CCT" at positions 2041-2043 mutated to "AGA". D950N (a single amino acid residue mutation) The "G" at position 2848 mutates to "A", and the "C" at position 2850 mutates to "T".

[0122] Table 6

[0123]

[0124]

[0125] Table 7

[0126] Protein mutation DNA mutation (corresponding to position 19 in sequence) S13I (a mutation involving one amino acid residue) The 38-bit "G" mutates to "T". W152C (a mutation involving one amino acid residue) The 456-bit "G" mutated to "C". L452R (a single amino acid residue mutation) The "T" at position 1355 mutates to "G". D614G (a single amino acid residue mutation) The "A" at position 1841 mutates to "G".

[0127] Table 8

[0128] Protein mutation DNA mutation (corresponding to position 19 in sequence) Q52R (a mutation involving one amino acid residue) The 155th position "A" mutates to "G". A67V (a mutation involving one amino acid residue) The 200th "C" mutates to a "T". 69-70del (missing two amino acid residues "HV") Missing bits 205-210 of "CACGTG" 144del (missing one amino acid residue "Y") The "TAT" character is missing in bits 430-432. E484K (a single amino acid residue mutation) The "G" at position 1450 mutated to "A". D614G (a single amino acid residue mutation) The "A" at position 1841 mutates to "G". Q677H (a mutation involving one amino acid residue) The "G" at position 2031 mutates to "C". F888L (a mutation involving one amino acid residue) The 2664th position "C" mutates to "A".

[0129] Table 9

[0130] Protein mutation DNA mutation (corresponding to position 19 in sequence) L5F (a mutation involving one amino acid residue) The 13th position "C" mutates to "T", and the 15th position "G" mutates to "C". T95I (a mutation involving one amino acid residue) The 284-bit "C" mutates to "T". D253G (a mutation involving one amino acid residue) The 758th bit "A" mutates to "G". E484K (a single amino acid residue mutation) The "G" at position 1450 mutated to "A". D614G (a single amino acid residue mutation) The "A" at position 1841 mutates to "G". A701V (a mutation involving one amino acid residue) The "C" at position 2102 mutates to "T".

[0131] Table 10

[0132] Protein mutation DNA mutation (corresponding to position 19 in sequence) N439K (a single amino acid residue mutation) The "C" at position 1350 mutated to "G".

[0133] II. Detection of Neutralizing Activity of Monoclonal Antibodies

[0134] 1. The test antibody was diluted using DMEM medium containing 10% FBS to obtain a diluted solution.

[0135] 2. Mix 100 μl of the diluent obtained in step 1 with 50 μl of the virus solution prepared in step 2 (virus content of 100 TCID50), and incubate at 37°C for 1 hour. Set up a blank control by replacing 100 μl of the diluent with 100 μl of DMEM medium containing 10% FBS.

[0136] 3. After completing step 2, add 50 μl of hACE2-hela cell suspension (containing approximately 2 × 10⁻⁶ cells). 4 (1 cell), incubated at 37°C for 48 hours.

[0137] 4. After completing step 3, add 100 μl of PBS buffer and 50 μl of cell lysis buffer (Bright-Globe). TM The Luciferase Assay System (Promega, E2650) was used to incubate the sample for 2 minutes, and then the luciferase activity was detected using a chemiluminescence analyzer.

[0138] Three replicates were set for each treatment, and the average value of the results was taken.

[0139] Neutralization activity = (fluorescence intensity of blank control group - fluorescence intensity of experimental group with added diluent) / fluorescence intensity of blank control group × 100%.

[0140] The antibody concentration corresponding to a neutralizing activity of 50% is the IC50 value.

[0141] The IC50 values ​​are shown in Tables 11 and 12. The units for the IC50 values ​​are μg / ml.

[0142] Six optimized antibodies (HX001-013, HX001-015, HX001-020, HX001-024, HX001-033, and HX001-034) exhibited strong neutralizing activity against wild-type SARS-CoV-2, as well as against various naturally occurring mutant strains. Compared to the P36-5D2 antibody, the neutralizing activity of the six optimized antibodies (HX001-013, HX001-015, HX001-020, HX001-024, HX001-033, and HX001-034) was increased by approximately 10-600 times.

[0143] Table 11

[0144] Wild-type novel coronavirus Alpha Beta Gamma Delta Delta Plus REGN10987 0.005 0.002 0.002 0.001 0.006 0.004 P36-5D2 0.053 0.025 0.039 0.032 2.651 2.230 HX001-013 0.008 0.002 0.006 0.003 0.007 0.006 HX001-015 0.006 0.003 0.003 0.003 0.022 0.024 HX001-020 0.008 0.004 0.002 0.003 0.008 0.004 HX001-024 0.005 0.004 0.002 0.005 0.006 0.003 HX001-033 0.006 0.005 0.002 0.002 0.019 0.011 HX001-034 0.015 0.012 0.009 0.008 0.023 0.014

[0145] Table 12

[0146] Kappa Eta Epsilon N439K Lota REGN10987 0.004 0.005 0.005 0.174 0.005 P36-5D2 1.795 1.139 4.323 1.059 1.474 HX001-013 0.011 0.011 0.004 0.007 0.010 HX001-015 0.030 0.007 0.024 0.004 0.004 HX001-020 0.009 0.009 0.006 0.005 0.008 HX001-024 0.010 0.010 0.006 0.005 0.012 HX001-033 0.021 0.006 0.017 0.006 0.004 HX001-034 0.027 0.020 0.028 0.010 0.026

[0147] The present invention has been described in detail above. For those skilled in the art, the invention can be practiced in a wide range of ways with equivalent parameters, concentrations, and conditions without departing from its spirit and scope, and without requiring unnecessary experiments. Although specific embodiments have been given, it should be understood that further modifications can be made to the invention. In summary, according to the principles of the invention, this application is intended to include any changes, uses, or improvements to the invention, including changes made using conventional techniques known in the art that depart from the scope disclosed herein. Some of the essential features can be applied within the scope of the following appended claims. sequence list <110> Tsinghua University Huashen Intelligent Pharmaceutical Technology (Beijing) Co., Ltd. <120> Six broad-spectrum neutralizing antibodies against SARS-CoV-2 and their applications <130> CGGNQYX216168 <160> twenty one <170> SIPOSequenceListing 1.0 <210> 1 <211> 233 <212> PRT <213> Artificial Sequence <400> 1 Met Gly Trp Ser Cys Ile Ile Leu Phe Leu Val Ala Thr Ala Thr Gly 1 5 10 15 Val His Ser Asp Ile Gln Met Thr Gln Ser Pro Ser Thr Leu Ser Ala 20 25 30 Ser Val Gly Asp Arg Val Thr Ile Thr Cys Arg Ala Ser Gln Ser Ile 35 40 45 Ser Ser Trp Leu Ala Trp Tyr Gln Gln Lys Pro Gly Lys Ala Pro Lys 50 55 60 Leu Leu Ile Tyr Asp Ala Ser Ser Leu Glu Ser Gly Val Pro Ser Arg 65 70 75 80 Phe Ser Gly Ser Gly Ser Gly Thr Glu Phe Thr Leu Thr Ile Ser Ser 85 90 95 Leu Gln Pro Asp Asp Phe Ala Thr Tyr Tyr Cys Gln Gln Tyr Asn Gly 100 105 110 Tyr Pro Trp Thr Phe Gly Gln Gly Thr Lys Val Glu Ile Lys Arg Thr 115 120 125 Val Ala Ala Pro Ser Val Phe Ile Phe Pro Pro Ser Asp Glu Gln Leu 130 135 140 Lys Ser Gly Thr Ala Ser Val Val Cys Leu Leu Asn Asn Phe Tyr Pro 145 150 155 160 Arg Glu Ala Lys Val Gln Trp Lys Val Asp Asn Ala Leu Gln Ser Gly 165 170 175 Asn Ser Gln Glu Ser Val Thr Glu Gln Asp Ser Lys Asp Ser Thr Tyr 180 185 190 Ser Leu Ser Ser Thr Leu Thr Leu Ser Lys Ala Asp Tyr Glu Lys His 195 200 205 Lys Val Tyr Ala Cys Glu Val Thr His Gln Gly Leu Ser Ser Pro Val 210 215 220 Thr Lys Ser Phe Asn Arg Gly Glu Cys 225 230 <210> 2 <211> 1887 <212> DNA <213> Artificial Sequence <400> 2 agtaatcaat tacggggtca ttagttcata gcccatatat ggagttccgc gttacataac 60 ttacggtaaa tggcccgcct ggctgaccgc ccaacgaccc ccgcccattg acgtcaataa 120 tgacgtatgt tcccatagta acgccaatag ggactttcca ttgacgtcaa tgggtggagt 180 atttacggta aactgcccac ttggcagtac atcaagtgta tcatatgcca agtacgcccc 240 ctattgacgt caatgacggt aaatggcccg cctggcatta tgcccagtac atgaccttat 300 gggactttcc tacttggcag tacatctacg tattagtcat cgctattacc atggtgatgc 360 ggttttggca gtacatcaat gggcgtggat agcggtttga ctcacgggga tttccaagtc 420 tccaccccat tgacgtcaat gggagtttgt tttggcacca aaatcaacgg gactttccaa 480 aatgtcgtaa caactccgcc ccattgacgc aaatgggcgg taggcgtgta cggtgggagg 540 tctatataag cagagctcgt ttagtgaacc gtcagatcgc ctggagacgc catccacgct 600 gttttgacct ccatagaaga caccgggacc gatccagcct ccatcggctc gcatctctcc 660 ttcacgcgcc cgccgcccta cctgaggccg ccatccacgc cggttgagtc gcgttctgcc 720 gcctcccgcc tgtggtgcct cctgaactgc gtccgccgtc taggtaagtt taaagctcag 780 gtcgagaccg ggcctttgtc cggcgctccc ttggagccta cctagactca gccggctctc 840 cacgctttgc ctgaccctgc ttgctcaact ctagttaacg gtggagggca gtgtagtctg 900 agcagtactc gttgctgccg cgcgcgccac cagacataat agctgacaga ctaacagact 960 gttcctttcc atgggtcttt tctgcagtca ccgtcgtcga cacgtgtgat cagatatcgc 1020 ggccgctcta gaccaccatg ggatggtcat gtatcatcct ttttctagta gcaactgcaa ccggtgtaca ttcagacatc cagatgaccc agtctccttc caccctgtct gcatctgtag gagacagagt caccatcact tgccgggcca gtcagagtat tagtagctgg ttggcctggt atcagcaga accagggaa gcccctaagc tcctgatcta tgatgcctcc agtttggaaa gtggggtccc atctaggttc agcggcagtg gatctgggac agaattcact ctcaccatca 1320. gcagcctgca gcctgatgat tttgcaactt attactgcca acagtataat ggttacccgt 1380 1440. ggacgttcgg ccaagggacc aaggtggaa tcaaacgtac ggtggctgca ccatctgtct tcatcttccc gccatctgat gagcagttga aatctggaac tgcctctgtt gtgtgcctgc tgaataactt ctaccccaga gaagccaaag tgcagtggaa ggtggacaac gccctgcaga gcggaaacag ccaggaaagc gtgacagagc aggattccaa ggattccaca tacagcctga gcagcacact gacactgtcc aaggccgact acgagaagca caaggtgtac gcctgcgaag tgacacacca gggactgtcc tcccctgtga caaagagctt caacagagga gaatgctgaa agcttggccg ccatggccca acttgtttat tgcagcttat aatggttaca aataaagcaa 1800 tagcatcaca aatttcacaa ataaagcatt tttttcactg cattctagtt gtggtttgtc 1860 caaactcatc aatgtatctt atcatgt 1887 <210> 3 <211> 471 <212> PRT <213> Artificial Sequence <400> 3 Met Gly Trp Ser Cys Ile Ile Leu Phe Leu Val Ala Thr Ala Thr Gly 1 5 10 15 Val His Ser Gln Val Gln Leu Val Gln Ser Gly Ala Glu Val Lys Lys 20 25 30 Pro Gly Ala Ser Val Lys Val Ser Cys Lys Ala Ser Gly Tyr Thr Phe 35 40 45 Thr Thr Tyr Ala Met His Trp Val Arg Gln Ala Pro Gly Gln Arg Leu 50 55 60 Glu Trp Met Gly Trp Ile Asn Ala Gly Asn Gly Asn Thr Lys Tyr Ser 65 70 75 80 Gln Lys Phe Gln Gly Arg Val Thr Ile Thr Arg Asp Thr Ser Ala Ser 85 90 95 Thr Ala Tyr Met Glu Leu Ser Ser Leu Arg Ser Glu Asp Thr Ala Val 100 105 110 Tyr Tyr Cys Ala Gly Gly Gly Gly Arg Met Phe Gln Phe Asp Tyr Phe 115 120 125 Asp Tyr Trp Gly Gln Gly Thr Leu Val Thr Val Ser Ser Ala Ser Thr 130 135 140 Lys Gly Pro Ser Val Phe Pro Leu Ala Pro Ser Ser Lys Ser Thr Ser 145 150 155 160 Gly Gly Thr Ala Ala Leu Gly Cys Leu Val Lys Asp Tyr Phe Pro Glu 165 170 175 Pro Val Thr Val Ser Trp Asn Ser Gly Ala Leu Thr Ser Gly Val His 180 185 190 Thr Phe Pro Ala Val Leu Gln Ser Ser Gly Leu Tyr Ser Leu Ser Ser 195 200 205 Val Val Thr Val Pro Ser Ser Ser Leu Gly Thr Gln Thr Tyr Ile Cys 210 215 220 Asn Val Asn His Lys Pro Ser Asn Thr Lys Val Asp Lys Arg Val Glu 225 230 235 240 Pro Lys Ser Cys Asp Lys Thr His Thr Cys Pro Pro Cys Pro Ala Pro 245 250 255 Glu Leu Leu Gly Gly Pro Ser Val Phe Leu Phe Pro Pro Lys Pro Lys 260 265 270 Asp Thr Leu Met Ile Ser Arg Thr Pro Glu Val Thr Cys Val Val Val 275 280 285 Asp Val Ser His Glu Asp Pro Glu Val Lys Phe Asn Trp Tyr Val Asp 290 295 300 Gly Val Glu Val His Asn Ala Lys Thr Lys Pro Arg Glu Glu Gln Tyr 305 310 315 320 Asn Ser Thr Tyr Arg Val Val Ser Val Leu Thr Val Leu His Gln Asp 325 330 335 Trp Leu Asn Gly Lys Glu Tyr Lys Cys Lys Val Ser Asn Lys Ala Leu 340 345 350 Pro Ala Pro Ile Glu Lys Thr Ile Ser Lys Ala Lys Gly Gln Pro Arg 355 360 365 Glu Pro Gln Val Tyr Thr Leu Pro Pro Ser Arg Glu Glu Met Thr Lys 370 375 380 Asn Gln Val Ser Leu Thr Cys Leu Val Lys Gly Phe Tyr Pro Ser Asp 385 390 395 400 Ile Ala Val Glu Trp Glu Ser Asn Gly Gln Pro Glu Asn Asn Tyr Lys 405 410 415 Thr Thr Pro Pro Val Leu Asp Ser Asp Gly Ser Phe Phe Leu Tyr Ser 420 425 430 Lys Leu Thr Val Asp Lys Ser Arg Trp Gln Gln Gly Asn Val Phe Ser 435 440 445 Cys Ser Val Met His Glu Ala Leu His Asn His Tyr Thr Gln Lys Ser 450 455 460 Leu Ser Leu Ser Pro Gly Lys 465 470 <210> 4 <211> 2453 <212> DNA <213> Artificial Sequence <400> 4 atagggactt tccattgacg tcaatgggtg gagtatttac ggtaaactgc ccacttggca 60 gtacatcaag tgtatcatat gccaagtacg ccccctattg acgtcaatga cggtaaatgg 120<00D0739>cccgcctggc attatgccca gtacatgacc ttatgggact ttcctacttg gcagtacatc 180 tacgtattag tcatcgctat taccatggtg atgcggtttt ggcagtacat caatgggcgt 240 ggatagcggt ttgactcacg gggatttcca agtctccacc ccattgacgt caatgggagt 300 ttgttttggc accaaaatca acgggacttt ccaaaatgtc gtaacaactc cgccccattg 360 It should be noted that there seems to be a small error in your original text where <00D0739> should probably be . This has been corrected in the translation.acgcaaatgg gcggtaggcg tgtacggtgg gaggtctata taagcagagc tcgtttagtg 420 aaccgtcaga tcgcctggag acgccatcca cgctgttttg acctccatag aagacaccgg 480 gaccgatcca gcctccatcg gctcgcatct ctccttcacg cgcccgccgc cctacctgag 540 gccgccatcc acgccggttg agtcgcgttc tgccgcctcc cgcctgtggt gcctcctgaa 600 ctgcgtccgc cgtctaggta agtttaaagc tcaggtcgag accgggcctt tgtccggcgc 660 tcccttggag cctacctaga ctcagccggc tctccacgct ttgcctgacc ctgcttgctc 720 aactctagtt aacggtggag ggcagtgtag tctgagcagt actcgttgct gccgcgcgcg 780 ccaccagaca taatagctga cagaccaca gactgttcct ttccatgggt cttttctgca 840 gtcaccgtcc tcgacacgtg tgatcagata tcgcggccgc tctagaccac catgggatgg 900 tcatgtatca tccttttct agtagcaact gcaaccggtg tacattccca ggtccagctt 960 gtgcagtctg gggctgaggt gaaagcct ggggcctcag tgaaggtttc ctgcaaggct 1020 tctggataca ccttcactac ctatgctatg cattgggtgc gccaggcccc cggacaaagg 1080 cttgagtgga tgggatggat caacgctggc aatggtaaca caaaatattc agaagttc 1140 cagggcagag tcaccattac caggagacaca tccgcgagca cagcctacat ggagctgagc 1200 agcctgagat ctgaagacac ggctgtgtat tactgtgcgg gaggggggg taggatgttc 1260 caatttgatt actttgacta ctggggccag ggaaccctgg tcaccgtctc ctcagcgtcg 1320 accaagggcc catcggtctt ccccctggca ccctcctcca agagcacctc tggggcaca 1380 gcggccctgg gctgcctggt caaggactac ttccccgaac ctgtgacggt ctcgtggaac 1440 tcaggcgcc tgaccagcgg cgtgcacacc ttcccggctg tcctacagtc ctcaggactc 1500 tactccctca gcagcgtggt gaccgtgccc tccagcagct tgggcaccca gacctacatc 1560 tgcaacgtga atcacaagcc cagcaacacc aaggtggaca agagagttga gcccaaatct 1620 tgtgacaaaa ctcacacatg cccaccgtgc ccagcacctg aactcctggg gggaccgtca 1680 gtttctctct tccccccaaa acccaaggac accctcatga tctcccggac ccctgaggtc 1740 acatgcgtgg tggtggacgt gagccacgaa gaccctgagg tcaagttcaa ctggtacgtg 1800 gacggcgtgg aggtgcataa tgccaagaca aagccgcggg aggagcagta caacagcacg 1860 taccgtgtgg tcagcgtcct caccgtcctg caccaggact ggctgaatgg caaggagtac 1920 aagtgcaagg tctccaacaa agccctccca gcccccatcg agaaaaccat ctccaaagcc 1980 aaagggcagc cccgagaacc acaggtgtac accctgcccc catcccggga ggagatgacc 2040 aagaaccagg tcagcctgac ctgcctggtc aaaggcttct atcccagcga catcgccgtg 2100 gagtgggaga gcaatgggca gccggagaac aactacaaga ccacgcctcc cgtgctggac 2160 tccgacggct ccttcttcct ctatagcaag ctcaccgtgg acaagagcag gtggcagcag 2220 gggaacgtct tctcatgctc cgtgatgcat gaggctctgc acaaccacta cacgcagaag 2280 agcctctccc tgtccccggg taaatgaaag cttggccgcc atggcccaac ttgtttattg 2340 cagcttataa tggttacaaa taaagcaata gcatcacaaa tttcacaaat aaagcatttt 2400 tttcactgca ttctagttgt ggtttgtcca aactcatcaa tgtatcttat cat 2453 <210> 5 <211> 471 <212> PRT <213> Artificial Sequence <400> 5 Met Gly Trp Ser Cys Ile Ile Leu Phe Leu Val Ala Thr Ala Thr Gly 1 5 10 15 Val His Ser Gln Val Gln Leu Val Gln Ser Gly Ala Glu Val Lys Lys 20 25 30 Pro Gly Ala Ser Val Lys Val Ser Cys Lys Ala Ser Gly Tyr Thr Phe 35 40 45 Thr Trp Tyr Ala Met His Trp Val Arg Gln Ala Pro Gly Gln Arg Leu 50 55 60 Glu Trp Met Gly Trp Ile Asn Ala Gly Asn Gly Asn Thr Lys Tyr Ser 65 70 75 80 Gln Lys Phe Gln Gly Arg Val Thr Ile Thr Arg Asp Thr Ser Ala Ser 85 90 95 Thr Ala Tyr Met Glu Leu Ser Ser Leu Arg Ser Glu Asp Thr Ala Val 100 105 110 Tyr Tyr Cys Ala Gly Gly Gly Gly Arg Met Leu Gln Phe Asp Tyr Phe 115 120 125 Asp Tyr Trp Gly Gln Gly Thr Leu Val Thr Val Ser Ser Ala Ser Thr 130 135 140 Lys Gly Pro Ser Val Phe Pro Leu Ala Pro Ser Ser Lys Ser Thr Ser 145 150 155 160 Gly Gly Thr Ala Ala Leu Gly Cys Leu Val Lys Asp Tyr Phe Pro Glu 165 170 175 Pro Val Thr Val Ser Trp Asn Ser Gly Ala Leu Thr Ser Gly Val His 180 185 190 Thr Phe Pro Ala Val Leu Gln Ser Ser Gly Leu Tyr Ser Leu Ser Ser 195 200 205 Val Val Thr Val Pro Ser Ser Ser Leu Gly Thr Gln Thr Tyr Ile Cys 210 215 220 Asn Val Asn His Lys Pro Ser Asn Thr Lys Val Asp Lys Arg Val Glu 225 230 235 240 Pro Lys Ser Cys Asp Lys Thr His Thr Cys Pro Pro Cys Pro Ala Pro 245 250 255 Glu Leu Leu Gly Gly Pro Ser Val Phe Leu Phe Pro Pro Lys Pro Lys 260 265 270 Asp Thr Leu Met Ile Ser Arg Thr Pro Glu Val Thr Cys Val Val Val 275 280 285 Asp Val Ser His Glu Asp Pro Glu Val Lys Phe Asn Trp Tyr Val Asp 290 295 300 Gly Val Glu Val His Asn Ala Lys Thr Lys Pro Arg Glu Glu Gln Tyr 305 310 315 320 Asn Ser Thr Tyr Arg Val Val Ser Val Leu Thr Val Leu His Gln Asp 325 330 335 Trp Leu Asn Gly Lys Glu Tyr Lys Cys Lys Val Ser Asn Lys Ala Leu 340 345 350 Pro Ala Pro Ile Glu Lys Thr Ile Ser Lys Ala Lys Gly Gln Pro Arg 355 360 365 Glu Pro Gln Val Tyr Thr Leu Pro Pro Ser Arg Glu Glu Met Thr Lys 370 375 380 Asn Gln Val Ser Leu Thr Cys Leu Val Lys Gly Phe Tyr Pro Ser Asp 385 390 395 400 Ile Ala Val Glu Trp Glu Ser Asn Gly Gln Pro Glu Asn Asn Tyr Lys 405 410 415 Thr Thr Pro Pro Val Leu Asp Ser Asp Gly Ser Phe Phe Leu Tyr Ser 420 425 430 Lys Leu Thr Val Asp Lys Ser Arg Trp Gln Gln Gly Asn Val Phe Ser 435 440 445 Cys Ser Val Met His Glu Ala Leu His Asn His Tyr Thr Gln Lys Ser 450 455 460 Leu Ser Leu Ser Pro Gly Lys 465 470 <210> 6 <211> 2453 <212> DNA <213> Artificial Sequence <400> 6 atagggactt tccattgacg tcaatgggtg gagtatttac ggtaaactgc ccacttggca 60 gtacatcaag tgtatcatat gccaagtacg ccccctattg acgtcaatga cggtaaatgg 120 cccgcctggc attatgccca gtacatgacc ttatgggact ttcctacttg gcagtacatc 180 tacgtattag tcatcgctat taccatggtg atgcggtttt ggcagtacat caatgggcgt 240 ggatagcggt ttgactcacg gggatttcca agtctccacc ccattgacgt caatgggagt 300 ttgttttggc accaaaatca acgggacttt ccaaaatgtc gtaacaactc cgccccattg 360 acgcaaatgg gcggtaggcg tgtacggtgg gaggtctata taagcagagc tcgtttagtg 420 aaccgtcaga tcgcctggag acgccatcca cgctgttttg acctccatag aagacaccgg 480 gaccgatcca gcctccatcg gctcgcatct ctccttcacg cgcccgccgc cctacctgag 540 gccgccatcc acgccggttg agtcgcgttc tgccgcctcc cgcctgtggt gcctcctgaa 600 ctgcgtccgc cgtctaggta agtttaaagc tcaggtcgag accgggcctt tgtccggcgc 660 tcccttggag cctacctaga ctcagccggc tctccacgct ttgcctgacc ctgcttgctc 720 aactctagtt aacggtggag ggcagtgtag tctgagcagt actcgttgct gccgcgcg 780 ccaccagaca tatagctga cagactaaca gactgttcct ttccatgggt cttttctgca 840 gtcaccgtcc tcgacacgtg tgatcagata tcgcggccgc tctagaccac catgggatgg 900 tcatgtatca tcctttttct agtagcaact gcaaccggtg tacattccca ggtccagctt 960 gtgcagtctg gggctgaggt gaagaagcct ggggcctcag tgaaggtttc ctgcaaggct 1020 tctggataca ccttcacttg gtatgctatg cattgggtgc gccaggcccc cggacaaagg 1080 cttgagtgga tgggatggat caacgctggc aatggtaaca caaaatattc acagaagttc 1140 cagggcagag tcaccattac cagggacaca tccgcgagca cagcctacat ggagctgagc 1200 agcctgagat ctgaagacac ggctgtgtat tactgtgcgg gagggggggg taggatgcta 1260 caatttgatt actttgacta ctggggccag ggaaccctgg tcaccgtctc ctcagcgtcg 1320 accaagggcc catcggtctt ccccctggca ccctcctcca agagcacctc tggggcaca 1380 gcggccctgg gctgcctggt caaggactac ttccccgaac ctgtgacggt ctcgtggaac 1440 tcaggcgcc tgaccagcgg cgtgcacacc ttcccggctg tcctacagtc ctcaggactc 1500 tactccctca gcagcgtggt gaccgtgccc tccagcagct tgggcaccca gacctacatc 1560 tgcaacgtga atcacaagcc cagcaacacc aaggtggaca agagagttga gcccaaatct 1620 tgtgacaaaa ctcacacatg cccaccgtgc ccagcacctg aactcctggg gggaccgtca 1680 gtttctctct tccccccaaa acccaaggac accctcatga tctcccggac ccctgaggtc 1740 acatgcgtgg tggtggacgt gagccacgaa gaccctgagg tcaagttcaa ctggtacgtg 1800 gacggcgtgg aggtgcataa tgccaagaca aagccgcggg aggagcagta caacagcacg 1860 taccgtgtgg tcagcgtcct caccgtcctg caccaggact ggctgaatgg caaggagtac 1920 aagtgcaagg tctccaacaa agccctccca gcccccatcg agaaaaccat ctccaaagcc 1980 aaagggcagc cccgagaacc acaggtgtac accctgcccc catcccggga ggagatgacc 2040 aagaaccagg tcagcctgac ctgcctggtc aaaggcttct atcccagcga catcgccgtg 2100 gagtgggaga gcaatgggca gccggagaac aactacaaga ccacgcctcc cgtgctggac 2160 tccgacggct ccttcttcct ctatagcaag ctcaccgtgg acaagagcag gtggcagcag 2220 gggaacgtct tctcatgctc cgtgatgcat gaggctctgc acaaccacta cacgcagaag 2280 agcctctccc tgtccccggg taaatgaaag cttggccgcc atggcccaac ttgtttattg 2340 cagcttataa tggttacaaa taaagcaata gcatcacaaa tttcacaaat aaagcatttt 2400 tttcactgca ttctagttgt ggtttgtcca aactcatcaa tgtatcttat cat 2453 <210> 7 <211> 471 <212> PRT <213> Artificial Sequence <400> 7 Met Gly Trp Ser Cys Ile Ile Leu Phe Leu Val Ala Thr Ala Thr Gly 1 5 10 15 Val His Ser Gln Val Gln Leu Val Gln Ser Gly Ala Glu Val Lys Lys 20 25 30 Pro Gly Ala Ser Val Lys Val Ser Cys Lys Ala Ser Gly Tyr Thr Phe 35 40 45 Thr Trp Tyr Ala Met His Trp Val Arg Gln Ala Pro Gly Gln Arg Leu 50 55 60 Glu Trp Met Gly Trp Ile Asn Ala Gly Asn Gly Asn Thr Lys Tyr Ser 65 70 75 80 Gln Lys Phe Gln Gly Arg Val Thr Ile Thr Arg Asp Thr Ser Ala Ser 85 90 95 Thr Ala Tyr Met Glu Leu Ser Ser Leu Arg Ser Glu Asp Thr Ala Val 100 105 110 Tyr Tyr Cys Ala Gly Gly Gly Gly Arg Met Phe Gln Phe Asp Tyr Phe 115 120 125 Asp Tyr Trp Gly Gln Gly Thr Leu Val Thr Val Ser Ser Ala Ser Thr 130 135 140 Lys Gly Pro Ser Val Phe Pro Leu Ala Pro Ser Ser Lys Ser Thr Ser 145 150 155 160 Gly Gly Thr Ala Ala Leu Gly Cys Leu Val Lys Asp Tyr Phe Pro Glu 165 170 175 Pro Val Thr Val Ser Trp Asn Ser Gly Ala Leu Thr Ser Gly Val His 180 185 190 Thr Phe Pro Ala Val Leu Gln Ser Ser Gly Leu Tyr Ser Leu Ser Ser 195 200 205 Val Val Thr Val Pro Ser Ser Ser Leu Gly Thr Gln Thr Tyr Ile Cys 210 215 220 Asn Val Asn His Lys Pro Ser Asn Thr Lys Val Asp Lys Arg Val Glu 225 230 235 240 Pro Lys Ser Cys Asp Lys Thr His Thr Cys Pro Pro Cys Pro Ala Pro 245 250 255 Glu Leu Leu Gly Gly Pro Ser Val Phe Leu Phe Pro Pro Lys Pro Lys 260 265 270 Asp Thr Leu Met Ile Ser Arg Thr Pro Glu Val Thr Cys Val Val Val 275 280 285 Asp Val Ser His Glu Asp Pro Glu Val Lys Phe Asn Trp Tyr Val Asp 290 295 300 Gly Val Glu Val His Asn Ala Lys Thr Lys Pro Arg Glu Glu Gln Tyr 305 310 315 320 Asn Ser Thr Tyr Arg Val Val Ser Val Leu Thr Val Leu His Gln Asp 325 330 335 Trp Leu Asn Gly Lys Glu Tyr Lys Cys Lys Val Ser Asn Lys Ala Leu 340 345 350 Pro Ala Pro Ile Glu Lys Thr Ile Ser Lys Ala Lys Gly Gln Pro Arg 355 360 365 Glu Pro Gln Val Tyr Thr Leu Pro Pro Ser Arg Glu Glu Met Thr Lys 370 375 380 Asn Gln Val Ser Leu Thr Cys Leu Val Lys Gly Phe Tyr Pro Ser Asp 385 390 395 400 Ile Ala Val Glu Trp Glu Ser Asn Gly Gln Pro Glu Asn Asn Tyr Lys 405 410 415 Thr Thr Pro Pro Val Leu Asp Ser Asp Gly Ser Phe Phe Leu Tyr Ser 420 425 430 Lys Leu Thr Val Asp Lys Ser Arg Trp Gln Gln Gly Asn Val Phe Ser 435 440 445 Cys Ser Val Met His Glu Ala Leu His Asn His Tyr Thr Gln Lys Ser 450 455 460 Leu Ser Leu Ser Pro Gly Lys 465 470 <210> 8 <211> 2453 <212> DNA <213> Artificial Sequence <400> 8 atagggactt tccattgacg tcaatgggtg gagtatttac ggtaaactgc ccacttggca 60 gtacatcaag tgtatcatat gccaagtacg ccccctattg acgtcaatga cggtaaatgg 120 It should be noted that there may be some errors in the original text you provided, such as the "420" in line 15 which might be a typo. I translated it as "420" as it was in the original. You may want to double-check the accuracy of the original text.cccgcctggc attatgccca gtacatgacc ttatgggact ttcctacttg gcagtacatc 180 tacgtattag tcatcgctat taccatggtg atgcggtttt ggcagtacat caatgggcgt 240 ggatagcggt ttgactcacg gggatttcca agtctccacc ccattgacgt caatgggagt 300 ttgttttggc accaaaatca acgggacttt ccaaaatgtc gtaacaactc cgccccattg 360 acgcaaatgg gcggtaggcg tgtacggtgg gaggtctata taagcagagc tcgtttagtg 420 aaccgtcaga tcgcctggag acgccatcca cgctgttttg acctccatag aagacaccgg 480 gaccgatcca gcctccatcg gctcgcatct ctccttcacg cgcccgccgc cctacctgag 540 gccgccatcc acgccggttg agtcgcgttc tgccgcctcc cgcctgtggt gcctcctgaa 600 ctgcgtccgc cgtctaggta agtttaaagc tcaggtcgag accgggcctt tgtccggcgc 660 tcccttggag cctacctaga ctcagccggc tctccacgct ttgcctgacc ctgcttgctc 720 aactctagtt aacggtggag ggcagtgtag tctgagcagt actcgttgct gccgcgcgcg 780 ccaccagaca taatagctga cagactaaca gactgttcct ttccatgggt cttttctgca 840 gtcaccgtcc tcgacacgtg tgatcagata tcgcggccgc tctagaccac catgggatgg 900 tcatgtatca tccttttct agtagcaact gcaaccggtg tacattccca ggtccagctt 960 gtgcagtctg gggctgaggt gaaagcct ggggcctcag tgaaggtttc ctgcaaggct 1020 tctggataca ccttcacttg gtatgctatg cattgggtgc gccaggcccc cggacaaagg 1080 cttgagtgga tgggatggat caacgctggc aatggtaaca caaaatattc agaagttc 1140 cagggcagag tcaccattac caggagacaca tccgcgagca cagcctacat ggagctgagc 1200 agcctgagat ctgaagacac ggctgtgtat tactgtgcgg gaggggggg taggatgttc 1260 caatttgatt actttgacta ctggggccag ggaaccctgg tcaccgtctc ctcagcgtcg 1320 accaagggcc catcggtctt ccccctggca ccctcctcca agagcacctc tggggcaca 1380 gcggccctgg gctgcctggt caaggactac ttccccgaac ctgtgacggt ctcgtggaac 1440 tcaggcgcc tgaccagcgg cgtgcacacc ttcccggctg tcctacagtc ctcaggactc 1500 tactccctca gcagcgtggt gaccgtgccc tccagcagct tgggcaccca gacctacatc 1560 tgcaacgtga atcacaagcc cagcaacacc aaggtggaca agagagttga gcccaaatct 1620 tgtgacaaaa ctcacacatg cccaccgtgc ccagcacctg aactcctggg gggaccgtca 1680 gtttctctct tccccccaaa acccaaggac accctcatga tctcccggac ccctgaggtc 1740 acatgcgtgg tggtggacgt gagccacgaa gaccctgagg tcaagttcaa ctggtacgtg 1800 gacggcgtgg aggtgcataa tgccaagaca aagccgcggg aggagcagta caacagcacg 1860 taccgtgtgg tcagcgtcct caccgtcctg caccaggact ggctgaatgg caaggagtac 1920 aagtgcaagg tctccaacaa agccctccca gcccccatcg agaaaaccat ctccaaagcc 1980 aaagggcagc cccgagaacc acaggtgtac accctgcccc catcccggga ggagatgacc 2040 aagaaccagg tcagcctgac ctgcctggtc aaaggcttct atcccagcga catcgccgtg 2100 gagtgggaga gcaatgggca gccggagaac aactacaaga ccacgcctcc cgtgctggac 2160 2220 gggaacgtct tctcatgctc cgtgatgcat gaggctctgc aaaccacta cacgcagaag 2280 agcctctccc tgtccccggg taaatgaaag cttggccgcc atggcccaac ttgtttattg 2340 cagcttataa tggttacaaa taaagcaata gcatcacaaa tttcacaaat aaagcatttt 2400 tttcactgca ttctagttgt ggtttgtcca aactcatcaa tgtatcttat cat 2453 <210> 9 <211> 471 <212> PRT <213> Artificial Sequence <400> 9 Met Gly Trp Ser Cys Ile Ile Leu Phe Leu Val Ala Thr Ala Thr Gly 1 5 10 15 Val His Ser Gln Val Gln Leu Val Gln Ser Gly Ala Glu Val Lys Lys 20 25 30 Pro Gly Ala Ser Val Lys Val Ser Cys Lys Ala Ser Gly Tyr Thr Phe 35 40 45 Thr Thr Tyr Ala Met His Trp Val Arg Gln Ala Pro Gly Gln Arg Leu 50 55 60 Glu Trp Met Gly Trp Ile Asn Ala Gly Asn Gly Leu Thr Lys Tyr Ser 65 70 75 80 Gln Lys Phe Gln Gly Arg Val Thr Ile Thr Arg Asp Thr Ser Ala Ser 85 90 95 Thr Ala Tyr Met Glu Leu Ser Ser Leu Arg Ser Glu Asp Thr Ala Val 100 105 110 Tyr Tyr Cys Ala Gly Gly Gly Gly Arg Met Phe Gln Phe Asp Tyr Phe 115 120 125 Asp Tyr Trp Gly Gln Gly Thr Leu Val Thr Val Ser Ser Ala Ser Thr 130 135 140 Lys Gly Pro Ser Val Phe Pro Leu Ala Pro Ser Ser Lys Ser Thr Ser 145 150 155 160 Gly Gly Thr Ala Ala Leu Gly Cys Leu Val Lys Asp Tyr Phe Pro Glu 165 170 175 Pro Val Thr Val Ser Trp Asn Ser Gly Ala Leu Thr Ser Gly Val His 180 185 190 Thr Phe Pro Ala Val Leu Gln Ser Ser Gly Leu Tyr Ser Leu Ser Ser 195 200 205 Val Val Thr Val Pro Ser Ser Ser Leu Gly Thr Gln Thr Tyr Ile Cys 210 215 220 Asn Val Asn His Lys Pro Ser Asn Thr Lys Val Asp Lys Arg Val Glu 225 230 235 240 Pro Lys Ser Cys Asp Lys Thr His Thr Cys Pro Pro Cys Pro Ala Pro 245 250 255 Glu Leu Leu Gly Gly Pro Ser Val Phe Leu Phe Pro Pro Lys Pro Lys 260 265 270 Asp Thr Leu Met Ile Ser Arg Thr Pro Glu Val Thr Cys Val Val Val 275 280 285 Asp Val Ser His Glu Asp Pro Glu Val Lys Phe Asn Trp Tyr Val Asp 290 295 300 Gly Val Glu Val His Asn Ala Lys Thr Lys Pro Arg Glu Glu Gln Tyr 305 310 315 320 Asn Ser Thr Tyr Arg Val Val Ser Val Leu Thr Val Leu His Gln Asp 325 330 335 Trp Leu Asn Gly Lys Glu Tyr Lys Cys Lys Val Ser Asn Lys Ala Leu 340 345 350 Pro Ala Pro Ile Glu Lys Thr Ile Ser Lys Ala Lys Gly Gln Pro Arg 355 360 365 Glu Pro Gln Val Tyr Thr Leu Pro Pro Ser Arg Glu Glu Met Thr Lys 370 375 380 Asn Gln Val Ser Leu Thr Cys Leu Val Lys Gly Phe Tyr Pro Ser Asp 385 390 395 400 Ile Ala Val Glu Trp Glu Ser Asn Gly Gln Pro Glu Asn Asn Tyr Lys 405 410 415 Thr Thr Pro Pro Val Leu Asp Ser Asp Gly Ser Phe Phe Leu Tyr Ser 420 425 430 Lys Leu Thr Val Asp Lys Ser Arg Trp Gln Gln Gly Asn Val Phe Ser 435 440 445 Cys Ser Val Met His Glu Ala Leu His Asn His Tyr Thr Gln Lys Ser 450 455 460 Leu Ser Leu Ser Pro Gly Lys 465 470 <210> 10 <211> 2453 <212> DNA <213> Artificial Sequence <400> 10 atagggactt tccattgacg tcaatgggtg gagtatttac ggtaaactgc ccacttggca 60 gtacatcaag tgtatcatat gccaagtacg ccccctattg acgtcaatga cggtaaatgg 120 cccgcctggc attatgccca gtacatgacc ttatgggact ttcctacttg gcagtacatc 180 tacgtattag tcatcgctat taccatggtg atgcggtttt ggcagtacat caatgggcgt 240 ggatagcggt ttgactcacg gggatttcca agtctccacc ccattgacgt caatgggagt 300 ttgttttggc accaaaatca acgggacttt ccaaaatgtc gtaacaactc cgccccattg 360 acgcaaatgg gcggtaggcg tgtacggtgg gaggtctata taagcagagc tcgtttagtg 420 aaccgtcaga tcgcctggag acgccatcca cgctgttttg acctccatag aagacaccgg 480 gaccgatcca gcctccatcg gctcgcatct ctccttcacg cgcccgccgc cctacctgag 540 gccgccatcc acgccggttg agtcgcgttc tgccgcctcc cgcctgtggt gcctcctgaa 600 ctgcgtccgc cgtctaggta agtttaaagc tcaggtcgag accgggcctt tgtccggcgc 660 tcccttggag cctacctaga ctcagccggc tctccacgct ttgcctgacc ctgcttgctc 720 aactctagtt aacggtggag ggcagtgtag tctgagcagt actcgttgct gccgcgcgcg 780 ccaccagaca taatagctga cagaccaca gactgttcct ttccatgggt cttttctgca 840 gtcaccgtcc tcgacacgtg tgatcagata tcgcggccgc tctagaccac catgggatgg 900 tcatgtatca tccttttct agtagcaact gcaaccggtg tacattccca ggtccagctt 960 gtgcagtctg gggctgaggt gaaagcct ggggcctcag tgaaggtttc ctgcaaggct 1020 tctggataca ccttcactac ctatgctatg cattgggtgc gccaggcccc cggacaaagg 1080 cttgagtgga tgggatggat caacgctggc aatggtctaa caaaatattc acagaagttc 1140 cagggcagag tcaccattac caggagacaca tccgcgagca cagcctacat ggagctgagc 1200 agcctgagat ctgaagacac ggctgtgtat tactgtgcgg gaggggggg taggatgttc 1260 caatttgatt actttgacta ctggggccag ggaaccctgg tcaccgtctc ctcagcgtcg 1320 accaagggcc catcggtctt ccccctggca ccctcctcca agagcacctc tggggcaca 1380 gcggccctgg gctgcctggt caaggactac ttccccgaac ctgtgacggt ctcgtggaac 1440 tcaggcgcc tgaccagcgg cgtgcacacc ttcccggctg tcctacagtc ctcaggactc 1500 tactccctca gcagcgtggt gaccgtgccc tccagcagct tgggcaccca gacctacatc 1560 tgcaacgtga atcacaagcc cagcaacacc aaggtggaca agagagttga gcccaaatct 1620 tgtgacaaaa ctcacacatg cccaccgtgc ccagcacctg aactcctggg gggaccgtca 1680 gtttctctct tccccccaaa acccaaggac accctcatga tctcccggac ccctgaggtc 1740 acatgcgtgg tggtggacgt gagccacgaa gaccctgagg tcaagttcaa ctggtacgtg 1800 gacggcgtgg aggtgcataa tgccaagaca aagccgcggg aggagcagta caacagcacg 1860 taccgtgtgg tcagcgtcct caccgtcctg caccaggact ggctgaatgg caaggagtac 1920 aagtgcaagg tctccaacaa agccctccca gcccccatcg agaaaaccat ctccaaagcc 1980 aaagggcagc cccgagaacc acaggtgtac accctgcccc catcccggga ggagatgacc 2040 aagaaccagg tcagcctgac ctgcctggtc aaaggcttct atcccagcga catcgccgtg 2100 gagtgggaga gcaatgggca gccggagaac aactacaaga ccacgcctcc cgtgctggac 2160 2220 gggaacgtct tctcatgctc cgtgatgcat gaggctctgc aaaccacta cacgcagaag 2280 agcctctccc tgtccccggg taaatgaaag cttggccgcc atggcccaac ttgtttattg 2340 cagcttataa tggttacaaa taaagcaata gcatcacaaa tttcacaaat aaagcatttt 2400 tttcactgca ttctagttgt ggtttgtcca aactcatcaa tgtatcttat cat 2453 <210> 11 <211> 471 <212> PRT <213> Artificial Sequence <400> 11 Met Gly Trp Ser Cys Ile Ile Leu Phe Leu Val Ala Thr Ala Thr Gly 1 5 10 15 Val His Ser Gln Val Gln Leu Val Gln Ser Gly Ala Glu Val Lys Lys 20 25 30 Pro Gly Ala Ser Val Lys Val Ser Cys Lys Ala Ser Gly Tyr Thr Phe 35 40 45 Thr Trp Tyr Ala Met His Trp Val Arg Gln Ala Pro Gly Gln Arg Leu 50 55 60 Glu Trp Met Gly Trp Ile Asn Ala Gly Asn Gly Leu Thr Lys Tyr Ser 65 70 75 80 Gln Lys Phe Gln Gly Arg Val Thr Ile Thr Arg Asp Thr Ser Ala Ser 85 90 95 Thr Ala Tyr Met Glu Leu Ser Ser Leu Arg Ser Glu Asp Thr Ala Val ]>100 105 110 Tyr Tyr Cys Ala Gly Gly Gly Gly Arg Met Leu Gln Phe Asp Tyr Phe 115 120 125 Asp Tyr Trp Gly Gln Gly Thr Leu Val Thr Val Ser Ser Ala Ser Thr[[ID=^37]] 130 135 140 Lys Gly Pro Ser Val Phe Pro Leu Ala Pro Ser Ser Lys Ser Thr Ser 145 150 155 160 Gly Gly Thr Ala Ala Leu Gly Cys Leu Val Lys Asp Tyr Phe Pro Glu 165 170 175 Pro Val Thr Val Ser Trp Asn Ser Gly Ala Leu Thr Ser Gly Val His 180 185 190 Thr Phe Pro Ala Val Leu Gln Ser Ser Gly Leu Tyr Ser Leu Ser Ser 195 200 205 Val Val Thr Val Pro Ser Ser Ser Leu Gly Thr Gln Thr Tyr Ile Cys 210 215 220 Asn Val Asn His Lys Pro Ser Asn Thr Lys Val Asp Lys Arg Val Glu 225 230 235 240 Pro Lys Ser Cys Asp Lys Thr His Thr Cys Pro Pro Cys Pro Ala Pro 245 250 255 Glu Leu Leu Gly Gly Pro Ser Val Phe Leu Phe Pro Pro Lys Pro Lys 260 265 270 Asp Thr Leu Met Ile Ser Arg Thr Pro Glu Val Thr Cys Val Val Val 275 280 285 Asp Val Ser His Glu Asp Pro Glu Val Lys Phe Asn Trp Tyr Val Asp 290 295 300 Gly Val Glu Val His Asn Ala Lys Thr Lys Pro Arg Glu Glu Gln Tyr 305 310 315 320 Asn Ser Thr Tyr Arg Val Val Ser Val Leu Thr Val Leu His Gln Asp 325 330 335 Trp Leu Asn Gly Lys Glu Tyr Lys Cys Lys Val Ser Asn Lys Ala Leu 340 345 350 Pro Ala Pro Ile Glu Lys Thr Ile Ser Lys Ala Lys Gly Gln Pro Arg 355 360 365 Glu Pro Gln Val Tyr Thr Leu Pro Pro Ser Arg Glu Glu Met Thr Lys 370 375 380 Asn Gln Val Ser Leu Thr Cys Leu Val Lys Gly Phe Tyr Pro Ser Asp 385 390 395 400 Ile Ala Val Glu Trp Glu Ser Asn Gly Gln Pro Glu Asn Asn Tyr Lys 405 410 415 Thr Thr Pro Pro Val Leu Asp Ser Asp Gly Ser Phe Phe Leu Tyr Ser 420 425 430 Lys Leu Thr Val Asp Lys Ser Arg Trp Gln Gln Gly Asn Val Phe Ser 435 440 445 Cys Ser Val Met His Glu Ala Leu His Asn His Tyr Thr Gln Lys Ser 450 455 460 Leu Ser Leu Ser Pro Gly Lys 465 470 <210> 12 <211> 2453 <212> DNA <213> Artificial Sequence <400> 12 atagggactt tccattgacg tcaatgggtg gagtatttac ggtaaactgc ccacttggca 60 gtacatcaag tgtatcatat gccaagtacg ccccctattg acgtcaatga cggtaaatgg 120 cccgcctggc attatgccca gtacatgacc ttatgggact ttcctacttg gcagtacatc 180 tacgtattag tcatcgctat taccatggtg atgcggtttt ggcagtacat caatgggcgt 240 ggatagcggt ttgactcacg gggatttcca agtctccacc ccattgacgt caatgggagt 300 ttgttttggc accaaaatca acgggacttt ccaaaatgtc gtaacaactc cgccccattg 360 acgcaaatgg gcggtaggcg tgtacggtgg gaggtctata taagcagagc tcgtttagtg 420 aaccgtcaga tcgcctggag acgccatcca cgctgttttg acctccatag aagacaccgg 480 gaccgatcca gcctccatcg gctcgcatct ctccttcacg cgcccgccgc cctacctgag 540 gccgccatcc acgccggttg agtcgcgtc tgccgcctcc cgcctgtggt gcctcctgaa 600 ctgcgtccgc cgtctaggta agtttaaagc tcaggtcgag accgggcctt tgtccggcgc 660 tcccttggag cctacctaga ctcagccggc tctccacgct ttgcctgacc ctgcttgctc 720 aactctagtt aacggtggag ggcagtgtag tctgagcagt actcgttgct gccgcgcg 780 ccaccagaca tatagctga cagactaaca gactgttcct ttccatgggt cttttctgca 840 gtcaccgtcc tcgacacgtg tgatcagata tcgcggccgc tctagaccac catgggatgg 900 tcatgtatca tcctttttct agtagcaact gcaaccggtg tacattccca ggtccagctt 960 gtgcagtctg gggctgaggt gaagaagcct ggggcctcag tgaaggtttc ctgcaaggct 1020 tctggataca ccttcacttg gtatgctatg cattgggtgc gccaggcccc cggacaaagg 1080 cttgagtgga tgggatggat caacgctggc aatggtctaa caaaatattc acagaagttc 1140 cagggcagag tcaccattac cagggacaca tccgcgagca cagcctacat ggagctgagc 1200 agcctgagat ctgaagacac ggctgtgtat tactgtgcgg gagggggggg taggatgcta 1260 caatttgatt actttgacta ctggggccag ggaaccctgg tcaccgtctc ctcagcgtcg 1320 accaagggcc catcggtctt ccccctggca ccctcctcca agagcacctc tggggcaca 1380 gcggccctgg gctgcctggt caaggactac ttccccgaac ctgtgacggt ctcgtggaac 1440 tcaggcgcc tgaccagcgg cgtgcacacc ttcccggctg tcctacagtc ctcaggactc 1500 tactccctca gcagcgtggt gaccgtgccc tccagcagct tgggcaccca gacctacatc 1560 tgcaacgtga atcacaagcc cagcaacacc aaggtggaca agagagttga gcccaaatct 1620 tgtgacaaaa ctcacacatg cccaccgtgc ccagcacctg aactcctggg gggaccgtca 1680 gtttctctct tccccccaaa acccaaggac accctcatga tctcccggac ccctgaggtc 1740 acatgcgtgg tggtggacgt gagccacgaa gaccctgagg tcaagttcaa ctggtacgtg 1800 gacggcgtgg aggtgcataa tgccaagaca aagccgcggg aggagcagta caacagcacg 1860 taccgtgtgg tcagcgtcct caccgtcctg caccaggact ggctgaatgg caaggagtac 1920 aagtgcaagg tctccaacaa agccctccca gcccccatcg agaaaaccat ctccaaagcc 1980 aaagggcagc cccgagaacc acaggtgtac accctgcccc catcccggga ggagatgacc 2040 aagaaccagg tcagcctgac ctgcctggtc aaaggcttct atcccagcga catcgccgtg 2100 gagtgggaga gcaatgggca gccggagaac aactacaaga ccacgcctcc cgtgctggac 2160 tccgacggct ccttcttcct ctatagcaag ctcaccgtgg acaagagcag gtggcagcag 2220 gggaacgtct tctcatgctc cgtgatgcat gaggctctgc acaaccacta cacgcagaag 2280 agcctctccc tgtccccggg taaatgaaag cttggccgcc atggcccaac ttgtttattg 2340 cagcttataa tggttacaaa taaagcaata gcatcacaaa tttcacaaat aaagcatttt 2400 tttcactgca ttctagttgt ggtttgtcca aactcatcaa tgtatcttat cat 2453 <210> 13 <211> 471 <212> PRT <213> Artificial Sequence <400> 13 Met Gly Trp Ser Cys Ile Ile Leu Phe Leu Val Ala Thr Ala Thr Gly 1 5 10 15 Val His Ser Gln Val Gln Leu Val Gln Ser Gly Ala Glu Val Lys Lys 20 25 30 Pro Gly Ala Ser Val Lys Val Ser Cys Lys Ala Ser Gly Tyr Thr Phe 35 40 45 Thr Trp Tyr Ala Met His Trp Val Arg Gln Ala Pro Gly Gln Arg Leu 50 55 60 Glu Trp Met Gly Trp Ile Asn Ala Gly Asn Gly Leu Thr Lys Tyr Ser 65 70 75 80 Gln Lys Phe Gln Gly Arg Val Thr Ile Thr Arg Asp Thr Ser Ala Ser 85 90 95 Thr Ala Tyr Met Glu Leu Ser Ser Leu Arg Ser Glu Asp Thr Ala Val 100 105 110 Tyr Tyr Cys Ala Gly Gly Gly Gly Arg Met Phe Gln Phe Asp Tyr Phe 115 120 125 Asp Tyr Trp Gly Gln Gly Thr Leu Val Thr Val Ser Ser Ala Ser Thr 130 135 140 Lys Gly Pro Ser Val Phe Pro Leu Ala Pro Ser Ser Lys Ser Thr Ser 145 150 155 160 Gly Gly Thr Ala Ala Leu Gly Cys Leu Val Lys Asp Tyr Phe Pro Glu 165 170 175 Pro Val Thr Val Ser Trp Asn Ser Gly Ala Leu Thr Ser Gly Val His 180 185 190 Thr Phe Pro Ala Val Leu Gln Ser Ser Gly Leu Tyr Ser Leu Ser Ser 195 200 205 Val Val Thr Val Pro Ser Ser Ser Leu Gly Thr Gln Thr Tyr Ile Cys 210 215 220 Asn Val Asn His Lys Pro Ser Asn Thr Lys Val Asp Lys Arg Val Glu 225 230 235 240 Pro Lys Ser Cys Asp Lys Thr His Thr Cys Pro Pro Cys Pro Ala Pro 245 250 255 Glu Leu Leu Gly Gly Pro Ser Val Phe Leu Phe Pro Pro Lys Pro Lys 260 265 270 Asp Thr Leu Met Ile Ser Arg Thr Pro Glu Val Thr Cys Val Val Val 275 280 285 Asp Val Ser His Glu Asp Pro Glu Val Lys Phe Asn Trp Tyr Val Asp 290 295 300 Gly Val Glu Val His Asn Ala Lys Thr Lys Pro Arg Glu Glu Gln Tyr 305 310 315 320 Asn Ser Thr Tyr Arg Val Val Ser Val Leu Thr Val Leu His Gln Asp 325 330 335 Trp Leu Asn Gly Lys Glu Tyr Lys Cys Lys Val Ser Asn Lys Ala Leu 340 345 350 Pro Ala Pro Ile Glu Lys Thr Ile Ser Lys Ala Lys Gly Gln Pro Arg 355 360 365 Glu Pro Gln Val Tyr Thr Leu Pro Pro Ser Arg Glu Glu Met Thr Lys 370 375 380 Asn Gln Val Ser Leu Thr Cys Leu Val Lys Gly Phe Tyr Pro Ser Asp 385 390 395 400 Ile Ala Val Glu Trp Glu Ser Asn Gly Gln Pro Glu Asn Asn Tyr Lys 405 410 415 Thr Thr Pro Pro Val Leu Asp Ser Asp Gly Ser Phe Phe Leu Tyr Ser 420 425 430 Lys Leu Thr Val Asp Lys Ser Arg Trp Gln Gln Gly Asn Val Phe Ser 435 440 445 Cys Ser Val Met His Glu Ala Leu His Asn His Tyr Thr Gln Lys Ser 450 455 460 Leu Ser Leu Ser Pro Gly Lys 465 470 <210> 14 <211> 2453 <212> DNA <213> Artificial Sequence <400> 14 atagggactt tccattgacg tcaatgggtg gagtatttac ggtaaactgc ccacttggca 60 gtacatcaag tgtatcatat gccaagtacg ccccctattg acgtcaatga cggtaaatgg 120 cccgcctggc attatgccca gtacatgacc ttatgggact ttcctacttg gcagtacatc 180 tacgtattag tcatcgctat taccatggtg atgcggtttt ggcagtacat caatgggcgt 240 ggatagcggt ttgactcacg gggatttcca agtctccacc ccattgacgt caatgggagt 300 ttgttttggc accaaaatca acgggacttt ccaaaatgtc gtaacaactc cgccccattg 360 acgcaaatgg gcggtaggcg tgtacggtgg gaggtctata taagcagagc tcgtttagtg 420 aaccgtcaga tcgcctggag acgccatcca cgctgttttg acctccatag aagacaccgg 480 gaccgatcca gcctccatcg gctcgcatct ctccttcacg cgcccgccgc cctacctgag 540 gccgccatcc acgccggttg agtcgcgttc tgccgcctcc cgcctgtggt gcctcctgaa 600 ctgcgtccgc cgtctaggta agtttaaagc tcaggtcgag accgggcctt tgtccggcgc 660 tcccttggag cctacctaga ctcagccggc tctccacgct ttgcctgacc ctgcttgctc 720 aactctagtt aacggtggag ggcagtgtag tctgagcagt actcgttgct gccgcgcgcg 780 ccaccagaca taatagctga cagaccaca gactgttcct ttccatgggt cttttctgca 840 gtcaccgtcc tcgacacgtg tgatcagata tcgcggccgc tctagaccac catgggatgg 900 tcatgtatca tccttttct agtagcaact gcaaccggtg tacattccca ggtccagctt 960 gtgcagtctg gggctgaggt gaaagcct ggggcctcag tgaaggtttc ctgcaaggct 1020 tctggataca ccttcacttg gtatgctatg cattgggtgc gccaggcccc cggacaaagg 1080 cttgagtgga tgggatggat caacgctggc aatggtctaa caaaatattc acagaagttc 1140 cagggcagag tcaccattac caggagacaca tccgcgagca cagcctacat ggagctgagc 1200 agcctgagat ctgaagacac ggctgtgtat tactgtgcgg gaggggggg taggatgttc 1260 caatttgatt actttgacta ctggggccag ggaaccctgg tcaccgtctc ctcagcgtcg 1320 accaagggcc catcggtctt ccccctggca ccctcctcca agagcacctc tggggcaca 1380 gcggccctgg gctgcctggt caaggactac ttccccgaac ctgtgacggt ctcgtggaac 1440 tcaggcgcc tgaccagcgg cgtgcacacc ttcccggctg tcctacagtc ctcaggactc 1500 tactccctca gcagcgtggt gaccgtgccc tccagcagct tgggcaccca gacctacatc 1560 tgcaacgtga atcacaagcc cagcaacacc aaggtggaca agagagttga gcccaaatct 1620 tgtgacaaaa ctcacacatg cccaccgtgc ccagcacctg aactcctggg gggaccgtca 1680 gtttctctct tccccccaaa acccaaggac accctcatga tctcccggac ccctgaggtc 1740 acatgcgtgg tggtggacgt gagccacgaa gaccctgagg tcaagttcaa ctggtacgtg 1800 gacggcgtgg aggtgcataa tgccaagaca aagccgcggg aggagcagta caacagcacg 1860 taccgtgtgg tcagcgtcct caccgtcctg caccaggact ggctgaatgg caaggagtac 1920 aagtgcaagg tctccaacaa agccctccca gcccccatcg agaaaaccat ctccaaagcc 1980 aaagggcagc cccgagaacc acaggtgtac accctgcccc catcccggga ggagatgacc 2040 aagaaccagg tcagcctgac ctgcctggtc aaaggcttct atcccagcga catcgccgtg 2100 gagtgggaga gcaatgggca gccggagaac aactacaaga ccacgcctcc cgtgctggac 2160 2220 gggaacgtct tctcatgctc cgtgatgcat gaggctctgc acaaccacta cacgcagaag 2280 agcctctccc tgtccccggg taaatgaaag cttggccgcc atggcccaac ttgtttattg 2340 cagcttataa tggttacaaa taaagcaata gcatcacaaa tttcacaaat aaagcatttt 2400 tttcactgca ttctagttgt ggtttgtcca aactcatcaa tgtatcttat cat 2453 <210> 15 <211> 471 <212> PRT <213> Artificial Sequence <400> 15 Met Gly Trp Ser Cys Ile Ile Leu Phe Leu Val Ala Thr Ala Thr Gly 1 5 10 15 Val His Ser Gln Val Gln Leu Val Gln Ser Gly Ala Glu Val Lys Lys 20 25 30 Pro Gly Ala Ser Val Lys Val Ser Cys Lys Ala Ser Gly Tyr Thr Phe 35 40 45 Thr Thr Tyr Ala Met His Trp Val Arg Gln Ala Pro Gly Gln Arg Leu 50 55 60 Glu Trp Met Gly Trp Ile Asn Ala Gly Asn Gly Asn Thr Lys Tyr Ser 65 70 75 80 Gln Lys Phe Gln Gly Arg Val Thr Ile Thr Arg Asp Thr Ser Ala Ser 85 90 95 Thr Ala Tyr Met Glu Leu Ser Ser Leu Arg Ser Glu Asp Thr Ala Val 100 105 110 Tyr Tyr Cys Ala Gly Gly Gly Gly Arg Arg Leu Gln Phe Asp Tyr Phe 115 120 125 Asp Tyr Trp Gly Gln Gly Thr Leu Val Thr Val Ser Ser Ala Ser Thr 130 135 140 Lys Gly Pro Ser Val Phe Pro Leu Ala Pro Ser Ser Lys Ser Thr Ser 145 150 155 160 Gly Gly Thr Ala Ala Leu Gly Cys Leu Val Lys Asp Tyr Phe Pro Glu 165 170 175 Pro Val Thr Val Ser Trp Asn Ser Gly Ala Leu Thr Ser Gly Val His 180 185 190 Thr Phe Pro Ala Val Leu Gln Ser Ser Gly Leu Tyr Ser Leu Ser Ser 195 200 205 Val Val Thr Val Pro Ser Ser Ser Leu Gly Thr Gln Thr Tyr Ile Cys 210 215 220 Asn Val Asn His Lys Pro Ser Asn Thr Lys Val Asp Lys Arg Val Glu 225 230 235 240 Pro Lys Ser Cys Asp Lys Thr His Thr Cys Pro Pro Cys Pro Ala Pro 245 250 255 Glu Leu Leu Gly Gly Pro Ser Val Phe Leu Phe Pro Pro Lys Pro Lys 260 265 270 Asp Thr Leu Met Ile Ser Arg Thr Pro Glu Val Thr Cys Val Val Val 275 280 285 Asp Val Ser His Glu Asp Pro Glu Val Lys Phe Asn Trp Tyr Val Asp 290 295 300 Gly Val Glu Val His Asn Ala Lys Thr Lys Pro Arg Glu Glu Gln Tyr 305 310 315 320 Asn Ser Thr Tyr Arg Val Val Ser Val Leu Thr Val Leu His Gln Asp 325 330 335 Trp Leu Asn Gly Lys Glu Tyr Lys Cys Lys Val Ser Asn Lys Ala Leu 340 345 350 Pro Ala Pro Ile Glu Lys Thr Ile Ser Lys Ala Lys Gly Gln Pro Arg 355 360 365 Glu Pro Gln Val Tyr Thr Leu Pro Pro Ser Arg Glu Glu Met Thr Lys 370 375 380 Asn Gln Val Ser Leu Thr Cys Leu Val Lys Gly Phe Tyr Pro Ser Asp 385 390 395 400 Ile Ala Val Glu Trp Glu Ser Asn Gly Gln Pro Glu Asn Asn Tyr Lys 405 410 415 Thr Thr Pro Pro Val Leu Asp Ser Asp Gly Ser Phe Phe Leu Tyr Ser 420 425 430 Lys Leu Thr Val Asp Lys Ser Arg Trp Gln Gln Gly Asn Val Phe Ser 435 440 445 Cys Ser Val Met His Glu Ala Leu His Asn His Tyr Thr Gln Lys Ser 450 455 460 Leu Ser Leu Ser Pro Gly Lys 465 470 <210> 16 <211> 2453 <212> DNA <213> Artificial Sequence <400> 16 atagggactt tccattgacg tcaatgggtg gagtatttac ggtaaactgc ccacttggca 60 gtacatcaag tgtatcatat gccaagtacg ccccctattg acgtcaatga cggtaaatgg 120 cccgcctggc attatgccca gtacatgacc ttatgggact ttcctacttg gcagtacatc 180 tacgtattag tcatcgctat taccatggtg atgcggtttt ggcagtacat caatgggcgt 240 ggatagcggt ttgactcacg gggatttcca agtctccacc ccattgacgt caatgggagt 300 ttgttttggc accaaaatca acgggacttt ccaaaatgtc gtaacaactc cgccccattg 360 acgcaaatgg gcggtaggcg tgtacggtgg gaggtctata taagcagagc tcgtttagtg 420 aaccgtcaga tcgcctggag acgccatcca cgctgttttg acctccatag aagacaccgg 480 gaccgatcca gcctccatcg gctcgcatct ctccttcacg cgcccgccgc cctacctgag 540 gccgccatcc acgccggttg agtcgcgttc tgccgcctcc cgcctgtggt gcctcctgaa 600 ctgcgtccgc cgtctaggta agtttaaagc tcaggtcgag accgggcctt tgtccggcgc 660 tcccttggag cctacctaga ctcagccggc tctccacgct ttgcctgacc ctgcttgctc 720 aactctagtt aacggtggag ggcagtgtag tctgagcagt actcgttgct gccgcgcgcg 780 ccaccagaca taatagctga cagactaaca gactgttcct ttccatgggt cttttctgca 840 gtcaccgtcc tcgacacgtg tgatcagata tcgcggccgc tctagaccac catgggatgg 900 tcatgtatca tcctttttct agtagcaact gcaaccggtg tacattccca ggtccagctt 960 gtgcagtctg gggctgaggt gaaagcct ggggcctcag tgaaggtttc ctgcaaggct 1020 tctggataca ccttcactac ctatgctatg cattgggtgc gccaggcccc cggacaaagg 1080 cttgagtgga tgggatggat caacgctggc aatggtaaca caaaatattc agaagttc 1140 cagggcagag tcaccattac caggagacaca tccgcgagca cagcctacat ggagctgagc 1200 agcctgagat ctgaagacac ggctgtgtat tactgtgcgg gaggggggg taggcggcta 1260 caatttgatt actttgacta ctggggccag ggaaccctgg tcaccgtctc ctcagcgtcg 1320 accaagggcc catcggtctt ccccctggca ccctcctcca agagcacctc tggggcaca 1380 gcggccctgg gctgcctggt caaggactac ttccccgaac ctgtgacggt ctcgtggaac 1440 tcaggcgcc tgaccagcgg cgtgcacacc ttcccggctg tcctacagtc ctcaggactc 1500 tactccctca gcagcgtggt gaccgtgccc tccagcagct tgggcaccca gacctacatc 1560 tgcaacgtga atcacaagcc cagcaacacc aaggtggaca agagagttga gcccaaatct 1620 tgtgacaaaa ctcacacatg cccaccgtgc ccagcacctg aactcctggg gggaccgtca 1680 gtttctctct tccccccaaa acccaaggac accctcatga tctcccggac ccctgaggtc 1740 acatgcgtgg tggtggacgt gagccacgaa gaccctgagg tcaagttcaa ctggtacgtg 1800 gacggcgtgg aggtgcataa tgccaagaca aagccgcggg aggagcagta caacagcacg 1860 taccgtgtgg tcagcgtcct caccgtcctg caccaggact ggctgaatgg caaggagtac 1920 aagtgcaagg tctccaacaa agccctccca gcccccatcg agaaaaccat ctccaaagcc 1980 aaagggcagc cccgagaacc acaggtgtac accctgcccc catcccggga ggagatgacc 2040 aagaaccagg tcagcctgac ctgcctggtc aaaggcttct atcccagcga catcgccgtg 2100 gagtgggaga gcaatgggca gccggagaac aactacaaga ccacgcctcc cgtgctggac 2160 2220 gggaacgtct tctcatgctc cgtgatgcat gaggctctgc aaaccacta cacgcagaag 2280 agcctctccc tgtccccggg taaatgaaag cttggccgcc atggcccaac ttgtttattg 2340 cagcttataa tggttacaaa taaagcaata gcatcacaaa tttcacaaat aaagcatttt 2400 tttcactgca ttctagttgt ggtttgtcca aactcatcaa tgtatcttat cat 2453 <210> 17 <211> 267 <212> PRT <213> Artificial Sequence <400> 17 Met Leu Leu Val Asn Gln Ser His Gln Gly Phe Asn Lys Glu His Thr 1 5 10 15 Ser Lys Met Val Ser Ala Ile Val Leu Tyr Val Leu Leu Ala Ala Ala 20 25 30 Ala His Ser Ala Phe Ala Arg Val Gln Pro Thr Glu Ser Ile Val Arg 35 40 45 Phe Pro Asn Ile Thr Asn Leu Cys Pro Phe Gly Glu Val Phe Asn Ala 50 55 60 Thr Arg Phe Ala Ser Val Tyr Ala Trp Asn Arg Lys Arg Ile Ser Asn 65 70 75 80 Cys Val Ala Asp Tyr Ser Val Leu Tyr Asn Ser Ala Ser Phe Ser Thr 85 90 95 Phe Lys Cys Tyr Gly Val Ser Pro Thr Lys Leu Asn Asp Leu Cys Phe 100 105 110 Thr Asn Val Tyr Ala Asp Ser Phe Val Ile Arg Gly Asp Glu Val Arg 115 120 125 Gln Ile Ala Pro Gly Gln Thr Gly Lys Ile Ala Asp Tyr Asn Tyr Lys 130 135 140 Leu Pro Asp Asp Phe Thr Gly Cys Val Ile Ala Trp Asn Ser Asn Asn 145 150 155 160 Leu Asp Ser Lys Val Gly Gly Asn Tyr Asn Tyr Leu Tyr Arg Leu Phe 165 170 175 Arg Lys Ser Asn Leu Lys Pro Phe Glu Arg Asp Ile Ser Thr Glu Ile 180 185 190 Tyr Gln Ala Gly Ser Thr Pro Cys Asn Gly Val Glu Gly Phe Asn Cys 195 200 205 Tyr Phe Pro Leu Gln Ser Tyr Gly Phe Gln Pro Thr Asn Gly Val Gly 210 215 220 Tyr Gln Pro Tyr Arg Val Val Val Leu Ser Phe Glu Leu Leu His Ala 225 230 235 240 Pro Ala Thr Val Cys Gly Pro Lys Lys Ser Thr Asn Leu Val Lys Asn 245 250 255 Lys Cys Val Asn Phe His His His His His His 260 265 <210> 18 <211> 1273 <212> PRT <213> SARS‑CoV‑2 <400> 18 Met Phe Val Phe Leu Val Leu Leu Pro Leu Val Ser Ser Gln Cys Val 1 5 10 15 Asn Leu Thr Thr Arg Thr Gln Leu Pro Pro Ala Tyr Thr Asn Ser Phe 20 25 30 Thr Arg Gly Val Tyr Tyr Pro Asp Lys Val Phe Arg Ser Ser Val Leu 35 40 45 His Ser Thr Gln Asp Leu Phe Leu Pro Phe Phe Ser Asn Val Thr Trp 50 55 60 Phe His Ala Ile His Val Ser Gly Thr Asn Gly Thr Lys Arg Phe Asp 65 70 75 80 Asn Pro Val Leu Pro Phe Asn Asp Gly Val Tyr Phe Ala Ser Thr Glu 85 90 95 Lys Ser Asn Ile Ile Arg Gly Trp Ile Phe Gly Thr Thr Leu Asp Ser 100 105 110 Lys Thr Gln Ser Leu Leu Ile Val Asn Asn Ala Thr Asn Val Val Ile 115 120 125 Lys Val Cys Glu Phe Gln Phe Cys Asn Asp Pro Phe Leu Gly Val Tyr 130 135 140 Tyr His Lys Asn Asn Lys Ser Trp Met Glu Ser Glu Phe Arg Val Tyr 145 150 155 160 Ser Ser Ala Asn Asn Cys Thr Phe Glu Tyr Val Ser Gln Pro Phe Leu 165 170 175 Met Asp Leu Glu Gly Lys Gln Gly Asn Phe Lys Asn Leu Arg Glu Phe 180 185 190 Val Phe Lys Asn Ile Asp Gly Tyr Phe Lys Ile Tyr Ser Lys His Thr 195 200 205 Pro Ile Asn Leu Val Arg Asp Leu Pro Gln Gly Phe Ser Ala Leu Glu 210 215 220 Pro Leu Val Asp Leu Pro Ile Gly Ile Asn Ile Thr Arg Phe Gln Thr 225 230 235 240 Leu Leu Ala Leu His Arg Ser Tyr Leu Thr Pro Gly Asp Ser Ser Ser 245 250 255 Gly Trp Thr Ala Gly Ala Ala Ala Tyr Tyr Val Gly Tyr Leu Gln Pro 260 265 270 Arg Thr Phe Leu Leu Lys Tyr Asn Glu Asn Gly Thr Ile Thr Asp Ala 275 280 285 Val Asp Cys Ala Leu Asp Pro Leu Ser Glu Thr Lys Cys Thr Leu Lys 290 295 300 Ser Phe Thr Val Glu Lys Gly Ile Tyr Gln Thr Ser Asn Phe Arg Val 305 310 315 320 Gln Pro Thr Glu Ser Ile Val Arg Phe Pro Asn Ile Thr Asn Leu Cys 325 330 335 Pro Phe Gly Glu Val Phe Asn Ala Thr Arg Phe Ala Ser Val Tyr Ala 340 345 350 Trp Asn Arg Lys Arg Ile Ser Asn Cys Val Ala Asp Tyr Ser Val Leu 355 360 365 Tyr Asn Ser Ala Ser Phe Ser Thr Phe Lys Cys Tyr Gly Val Ser Pro 370 375 380 Thr Lys Leu Asn Asp Leu Cys Phe Thr Asn Val Tyr Ala Asp Ser Phe 385 390 395 400 Val Ile Arg Gly Asp Glu Val Arg Gln Ile Ala Pro Gly Gln Thr Gly 405 410 415 Lys Ile Ala Asp Tyr Asn Tyr Lys Leu Pro Asp Asp Phe Thr Gly Cys 420 425 430 Val Ile Ala Trp Asn Ser Asn Asn Leu Asp Ser Lys Val Gly Gly Asn 435 440 445 Tyr Asn Tyr Leu Tyr Arg Leu Phe Arg Lys Ser Asn Leu Lys Pro Phe 450 455 460 Glu Arg Asp Ile Ser Thr Glu Ile Tyr Gln Ala Gly Ser Thr Pro Cys 465 470 475 480 Asn Gly Val Glu Gly Phe Asn Cys Tyr Phe Pro Leu Gln Ser Tyr Gly 485 490 495 Phe Gln Pro Thr Asn Gly Val Gly Tyr Gln Pro Tyr Arg Val Val Val 500 505 510 Leu Ser Phe Glu Leu Leu His Ala Pro Ala Thr Val Cys Gly Pro Lys 515 520 525 Lys Ser Thr Asn Leu Val Lys Asn Lys Cys Val Asn Phe Asn Phe Asn 530 535 540 Gly Leu Thr Gly Thr Gly Val Leu Thr Glu Ser Asn Lys Lys Phe Leu 545 550 555 560 Pro Phe Gln Gln Phe Gly Arg Asp Ile Ala Asp Thr Thr Asp Ala Val 565 570 575 Arg Asp Pro Gln Thr Leu Glu Ile Leu Asp Ile Thr Pro Cys Ser Phe 580 585 590 Gly Gly Val Ser Val Ile Thr Pro Gly Thr Asn Thr Ser Asn Gln Val 595 600 605 Ala Val Leu Tyr Gln Asp Val Asn Cys Thr Glu Val Pro Val Ala Ile 610 615 620 His Ala Asp Gln Leu Thr Pro Thr Trp Arg Val Tyr Ser Thr Gly Ser 625 630 635 640 Asn Val Phe Gln Thr Arg Ala Gly Cys Leu Ile Gly Ala Glu His Val 645 650 655 Asn Asn Ser Tyr Glu Cys Asp Ile Pro Ile Gly Ala Gly Ile Cys Ala 660 665 670 Ser Tyr Gln Thr Gln Thr Asn Ser Pro Arg Arg Ala Arg Ser Val Ala 675 680 685 Ser Gln Ser Ile Ile Ala Tyr Thr Met Ser Leu Gly Ala Glu Asn Ser 690 695 700 Val Ala Tyr Ser Asn Asn Ser Ile Ala Ile Pro Thr Asn Phe Thr Ile 705 710 715 720 Ser Val Thr Thr Glu Ile Leu Pro Val Ser Met Thr Lys Thr Ser Val 725 730 735 Asp Cys Thr Met Tyr Ile Cys Gly Asp Ser Thr Glu Cys Ser Asn Leu 740 745 750 Leu Leu Gln Tyr Gly Ser Phe Cys Thr Gln Leu Asn Arg Ala Leu Thr 755 760 765 Gly Ile Ala Val Glu Gln Asp Lys Asn Thr Gln Glu Val Phe Ala Gln 770 775 780 Val Lys Gln Ile Tyr Lys Thr Pro Pro Ile Lys Asp Phe Gly Gly Phe 785 790 795 800 Asn Phe Ser Gln Ile Leu Pro Asp Pro Ser Lys Pro Ser Lys Arg Ser 805 810 815 Phe Ile Glu Asp Leu Leu Phe Asn Lys Val Thr Leu Ala Asp Ala Gly 820 825 830 Phe Ile Lys Gln Tyr Gly Asp Cys Leu Gly Asp Ile Ala Ala Arg Asp 835 840 845 Leu Ile Cys Ala Gln Lys Phe Asn Gly Leu Thr Val Leu Pro Pro Leu 850 855 860 Leu Thr Asp Glu Met Ile Ala Gln Tyr Thr Ser Ala Leu Leu Ala Gly 865 870 875 880 Thr Ile Thr Ser Gly Trp Thr Phe Gly Ala Gly Ala Ala Leu Gln Ile 885 890 895 Pro Phe Ala Met Gln Met Ala Tyr Arg Phe Asn Gly Ile Gly Val Thr 900 905 910 Gln Asn Val Leu Tyr Glu Asn Gln Lys Leu Ile Ala Asn Gln Phe Asn 915 920 925 Ser Ala Ile Gly Lys Ile Gln Asp Ser Leu Ser Ser Thr Ala Ser Ala 930 935 940 Leu Gly Lys Leu Gln Asp Val Val Asn Gln Asn Ala Gln Ala Leu Asn 945 950 955 960 Thr Leu Val Lys Gln Leu Ser Ser Asn Phe Gly Ala Ile Ser Ser Val 965 970 975 Leu Asn Asp Ile Leu Ser Arg Leu Asp Lys Val Glu Ala Glu Val Gln 980 985 990 Ile Asp Arg Leu Ile Thr Gly Arg Leu Gln Ser Leu Gln Thr Tyr Val 995 1000 1005 Thr Gln Gln Leu Ile Arg Ala Ala Glu Ile Arg Ala Ser Ala Asn Leu 1010 1015 1020 Ala Ala Thr Lys Met Ser Glu Cys Val Leu Gly Gln Ser Lys Arg Val 1025 1030 1035 1040 Asp Phe Cys Gly Lys Gly Tyr His Leu Met Ser Phe Pro Gln Ser Ala 1045 1050 1055 Pro His Gly Val Val Phe Leu His Val Thr Tyr Val Pro Ala Gln Glu 1060 1065 1070 Lys Asn Phe Thr Thr Ala Pro Ala Ile Cys His Asp Gly Lys Ala His 1075 1080 1085 Phe Pro Arg Glu Gly Val Phe Val Ser Asn Gly Thr His Trp Phe Val 1090 1095 1100 Thr Gln Arg Asn Phe Tyr Glu Pro Gln Ile Ile Thr Thr Asp Asn Thr 1105 1110 1115 1120 Phe Val Ser Gly Asn Cys Asp Val Val Ile Gly Ile Val Asn Asn Thr 1125 1130 1135 Val Tyr Asp Pro Leu Gln Pro Glu Leu Asp Ser Phe Lys Glu Glu Leu 1140 1145 1150 Asp Lys Tyr Phe Lys Asn His Thr Ser Pro Asp Val Asp Leu Gly Asp 1155 1160 1165 Ile Ser Gly Ile Asn Ala Ser Val Val Asn Ile Gln Lys Glu Ile Asp 1170 1175 1180 Arg Leu Asn Glu Val Ala Lys Asn Leu Asn Glu Ser Leu Ile Asp Leu 1185 1190 1195 1200 Gln Glu Leu Gly Lys Tyr Glu Gln Tyr Ile Lys Trp Pro Trp Tyr Ile 1205 1210 1215 Trp Leu Gly Phe Ile Ala Gly Leu Ile Ala Ile Val Met Val Thr Ile 1220 1225 1230 Met Leu Cys Cys Met Thr Ser Cys Cys Ser Cys Leu Lys Gly Cys Cys 1235 1240 1245 Ser Cys Gly Ser Cys Cys Lys Phe Asp Glu Asp Asp Ser Glu Pro Val 1250 1255 1260 Leu Lys Gly Val Lys Leu His Tyr Thr 1265 1270 <210> 19 <211> 3822 <212> DNA <213> SARS-CoV-2 <400> 19 atgttcgtgt tcctggtgct gctgcctctg gtgagcagcc agtgcgtgaa tctgaccacc 60 agaacccagc tgcctcctgc ctacaccaat agcttcacca gaggagttta ttatcccgat 120 aaggtgttca gaagtagtgt attacatagt acccaggacc tgttcctacc tttcttcagt 180 aacgtgacct ggttccacgc catccacgtg agcggcacca atggcaccaa gagattcgac 240 aatcctgtgc tgcctttcaa tgacggcgtg tacttcgcca gcaccgagaa gagcaatatc 300 atcagaggct ggatcttcgg caccaccttg gattccaaga ctcagagcct gctgattgta 360 aacaacgcta caaatgtggt gatcaaggtg tgcgagttcc agttctgcaa tgaccctttc 420 ctgggtgttt attatcataa gaacaacaag agctggatgg agagcgagtt ccgcgtatat 480 tcgtcggcta ataattgcac cttcgagtac gtgagccagc ctttcctgat ggacctggag 540 ggcaagcagg gcaatttcaa gaatctgaga gagttcgtgt tcaagaatat cgacggctac 600 ttcaagatct acagcaagca cacacccatt aatctggtga gagacctgcc tcagggcttc 660 agcgccctgg agcctctggt ggacctgcct atcggcatca atatcaccag attccagacc 720 ctgctggccc tgcacagatc atatcttaca ccaggcgatt cgtcaagcgg ttggaccgct 780 ggagctgcgg catattacgt gggctacctg cagcctagaa ccttcctgct gaagtacaat 840 gagaatggta cgataaccga cgcagttgat tgtgccctgg accctctgag cgagaccaag 900 tgcaccctga agagcttcac cgtggagaag ggcatctacc agaccagcaa tttcagagtg 960 cagcctaccg agagcatcgt gagattccct aatatcacca atctgtgccc ttcggcgag 1020 gtgttcaatg ccaccagatt cgccagcgtg tacgcatgga accgcaagcg gataagcaat 1080 tgcgtggccg actacagcgt gctgtacaat agcgccagct tcagcacctt caaatgttat 1140 ggtgttcgc caacaaagct gaatgacctg tgcttcacca atgtgtacgc cgacagcttc 1200 gtgatcagag gcgacgaggt gagacagatc gcgccagggc agaccggcaa gatcgccgac 1260 1320 1380 ctgaagcctt tcgagagaga catcagcacc gagatctacc aggccggcag cacaccgtgt 1440 aatggcgtgg agggcttcaa ttgctacttc cctctgcaga gctacggctt ccagcctacc 1500 aatggcgtgg gctaccagcc ttacagagtg gtggtgctga gcttcgagct gctccacgct 1560 cccgctaccg tgtgcggccc taagaagc accaatctgg tgaagaataa gtgcgtgaat 1620 ttcaatttca atggtctaac tggaacgggc gtgctgaccg agagcaataa gaagtttctt 1680 ccctttcaac aattcggcag agacatcgcc gacaccacag atgctgtaag agaccctcag 1740 accctggaga tcctggacat cactccgtgt agcttcggcg gcgtgagcgt gatcacaccg 1800 ggtaccaata ccagcaatca ggtggccgtg ctgtaccagg acgtgaattg caccgaggtg 1860 cctgtggcca tccacgccga ccagctgact cccacttgga gggtatattc cacgggaagc 1920 aatgtgttcc agaccagagc cggctgcctg atcggcgccg agcacgtgaa tatagctac 1980 gagtgcgaca tccctatcgg cgccggcatc tgcgccagct accagaccca gaccaatagc 2040 cctagaagag ccagaagcgt ggccagccag agcatcatcg cctacaccat gagcctgggc 2100 gccgagaata gcgtggccta cagcaataat agcatcgcca tccctaccaa tttcaccatc 2160 agcgtgacca ccgaaatatt accagtctcc atgaccaaga ccagcgtgga ctgcaccatg 2220 tacatctgcg gcgacagcac cgagtgcagc aatctgctgc tgcagtacgg cagcttctgc 2280 acccagctga atagagccct gaccggcatc gccgtggagc aggacaagaa tacccaggag 2340 gtgttcgccc aggtgaagca gatctacaag actccgccga tcaaggactt cggcggcttc 2400 aatttcagcc aaatactccc agatccaagc aagcctagca agaggagctt catcgaggac 2460 ctgctgttca ataaggtgac cctggccgac gccggcttca tcaagcagta cggcgactgc 2520 ctaggtgata ttgcggcaag agacctgatc tgcgcccaga agtttaacgg tttgacagta 2580 ctacctcctc tgctgaccga cgagatgata gcacaatata cgtcggcatt gctcgctggc 2640 acgatcacat cgggctggac tttcggcgcc ggagcagcgt tgcaaatccc tttcgccatg 2700 cagatggcct acagattcaa tggcatcggc gtgacccaga atgtgctgta cgagaatcag 2760 aagctgatcg ccaatcagtt caatagcgcc atcggcaaga tccaggacag cctgagcagc 2820 accgccagcg ccctgggcaa gctgcaggac gtggtgaatc agaatgccca ggccctgaat 2880 accctggtga agcagctgag cagcaatttc ggcgccatca gtagtgtact caacgatatc 2940 ctgagcagac tggacaaggt ggaggccgag gtgcaaattg atcgtcttat tacggcaga 3000 ctgcagagcc tgcagaccta cgtgacccag cagctgatca gagccgccga gatcagagcc 3060 agcgccaatc tggccgccac caagatgagc gagtgcgtgc tgggccagag caagagagtg 3120 gacttctgcg gcaagggcta ccacctgatg agcttccctc agagcgctcc acatggcgtg 3180 gtgttcctgc acgtgaccta cgtgcctgcc your foot atttcaccac cgcacccgca 3240 atctgccacg acggcaaggc ccacttccct agagagggcg tgttcgtgag caatggcacc 3300 cactggttcg tgacccagag aaatttctac gagcctcaga tcatcaccac cgacaatacc 3360 3420 ctgcagcctg agctggacag cttcaaggag gagctggaca agtacttcaa gaatcacacc 3480 agccctgacg tggacctcgg tgatatttcg ggaatcaatg ccagcgtggt gaatatccag 3540 aaggaaattg atcggctcaa cgaagtggcc aagaatctga atgagagcct gatcgacctg 3600 caggagctgg gcaagtacga gcagtacatc aagtggcctt ggtacatctg gctggggcttc 3660 atcgccggcc tgatcgccat cgtgatggtg accatcatgc tgtgctgcat gacctcctgt 3720 tgttcctgtt tgaaagggtg ttgttcgtgt gggtcctgct gcaagttcga cgaggacgac 3780 agcgagcctg tgctgaaggg cgtgaagctg cactacacct ga 3822 <210> 20 <211> 225 <212> PRT <213> Artificial Sequence <400> 20 Gln Val Gln Leu Val Glu Ser Gly Gly Gly Val Val Gln Pro Gly Arg 1 5 10 15 Ser Leu Arg Leu Ser Cys Ala Ala Ser Gly Phe Thr Phe Ser Asn Tyr 20 25 30 Ala Met Tyr Trp Val Arg Gln Ala Pro Gly Lys Gly Leu Glu Trp Val 35 40 45 Ala Val Ile Ser Tyr Asp Gly Ser Asn Lys Tyr Tyr Ala Asp Ser Val 50 55 60 Lys Gly Arg Phe Thr Ile Ser Arg Asp Asn Ser Lys Asn Thr Leu Tyr 65 70 75 80 Leu Gln Met Asn Ser Leu Arg Thr Glu Asp Thr Ala Val Tyr Tyr Cys 85 90 95 Ala Ser Gly Ser Asp Tyr Gly Asp Tyr Leu Leu Val Tyr Trp Gly Gln 100 105 110 Gly Thr Leu Val Thr Val Ser Ser Ala Ser Thr Lys Gly Pro Ser Val 115 120 125 Phe Pro Leu Ala Pro Ser Ser Lys Ser Thr Ser Gly Gly Thr Ala Ala 130 135 140 Leu Gly Cys Leu Val Lys Asp Tyr Phe Pro Glu Pro Val Thr Val Ser 145 150 155 160 Trp Asn Ser Gly Ala Leu Thr Ser Gly Val His Thr Phe Pro Ala Val 165 170 175 Leu Gln Ser Ser Gly Leu Tyr Ser Leu Ser Ser Val Val Thr Val Pro 180 185 190 Ser Ser Ser Leu Gly Thr Gln Thr Tyr Ile Cys Asn Val Asn His Lys 195 200 205 Pro Ser Asn Thr Lys Val Asp Lys Lys Val Glu Pro Lys Ser Cys Asp 210 215 220 Lys 225 <210> 21 <211> 216 <212> PRT <213> Artificial Sequence <400> 21 Gln Ser Ala Leu Thr Gln Pro Ala Ser Val Ser Gly Ser Pro Gly Gln 1 5 10 15 Ser Ile Thr Ile Ser Cys Thr Gly Thr Ser Ser Asp Val Gly Gly Tyr 20 25 30 Asn Tyr Val Ser Trp Tyr Gln Gln His Pro Gly Lys Ala Pro Lys Leu 35 40 45 Met Ile Tyr Asp Val Ser Lys Arg Pro Ser Gly Val Ser Asn Arg Phe 50 55 60 Ser Gly Ser Lys Ser Gly Asn Thr Ala Ser Leu Thr Ile Ser Gly Leu 65 70 75 80 Gln Ser Glu Asp Glu Ala Asp Tyr Tyr Cys Asn Ser Leu Thr Ser Ile 85 90 95 Ser Thr Trp Val Phe Gly Gly Gly Thr Lys Leu Thr Val Leu Gly Gln 100 105 110 Pro Lys Ala Ala Pro Ser Val Thr Leu Phe Pro Pro Ser Ser Glu Glu 115 120 125 Leu Gln Ala Asn Lys Ala Thr Leu Val Cys Leu Ile Ser Asp Phe Tyr 130 135 140 Pro Gly Ala Val Thr Val Ala Trp Lys Ala Asp Ser Ser Pro Val Lys 145 150 155 160 Ala Gly Val Glu Thr Thr Thr Pro Ser Lys Gln Ser Asn Asn Lys Tyr 165 170 175 Ala Ala Ser Ser Tyr Leu Ser Leu Thr Pro Glu Gln Trp Lys Ser His 180 185 190 Arg Ser Tyr Ser Cys Gln Val Thr His Glu Gly Ser Thr Val Glu Lys 195 200 205 Thr Val Ala Pro Thr Glu Cys Ser 210 215

Claims

1. An IgG antibody that binds SARS-CoV-2, characterized in that: The IgG antibody is composed of a light chain and a heavy chain; the CDR1, CDR2 and CDR3 in the variable region of the heavy chain are amino acid residues from the N-terminus of SEQ ID NO: 7, positions 45-52, 70-77 and 116-130 respectively; the CDR1, CDR2 and CDR3 in the variable region of the light chain are amino acid residues from the N-terminus of SEQ ID NO: 1, positions 46-51, 69-71 and 108-116 respectively.

2. The IgG antibody of claim 1, wherein: The heavy chain variable region is composed of amino acid residues from the N-terminus 20 to 142 of SEQ ID NO: 7; the light chain variable region is composed of amino acid residues from the N-terminus 20 to 126 of SEQ ID NO:

1.

3. The IgG antibody as described in claim 2, characterized in that: The heavy chain is shown in SEQ ID NO: 7; the light chain is shown in SEQ ID NO:

1.

4. A gene encoding the IgG antibody of any one of claims 1 to 3.

5. The use of the IgG antibody according to claim 1, 2 or 3 in the preparation of a medicament for neutralizing the novel coronavirus.

6. A drug for neutralizing the novel coronavirus, wherein the active ingredient is the IgG antibody as described in claim 1, 2 or 3.

7. The use of the IgG antibody according to claim 1, 2 or 3 in the preparation of a medicament for treating COVID-19.

8. A medicament for treating COVID-19, wherein the active ingredient is the IgG antibody as described in claim 1, 2 or 3.

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