Norovirus virus-like particles and uses thereof

Abstract

The application provides a norovirus virus-like particle and purposes thereof, the virus-like particle or an active fragment thereof comprising or consisting of an amino acid sequence shown in SEQ ID NO: 5. The application also provides a composition or kit comprising the norovirus virus-like particle and purposes of the norovirus virus-like particle. The single antigen of the application achieves the purpose of covering two valence types, can reduce the process difficulty of the preparation of a pharmaceutical composition or a vaccine, reduce the production cost, and has a broad market prospect.

Description

Technical Field

[0001] This invention belongs to the field of biopharmaceuticals and relates to a norovirus virus-like particle. This invention also provides the use of the said norovirus virus-like particle. Background Technology

[0002] Norovirus was first discovered and named Norwalk virus (NoV) in 1972 by American scholar Kapikian in the feces of diarrhea patients in Norwalk. It was subsequently named Norovirus at the 8th International Committee on Virus Nomenclature. It is one of the main pathogens causing outbreaks of infectious gastroenteritis in humans and acute diarrhea in infants and young children.

[0003] Norovirus is a single-stranded, positive-sense RNA virus belonging to the Caliciviridae family. It is approximately 26-35 nm in diameter, non-enveloped, rough-surfaced, spherical, and icosahedral in shape. It is isolated from the feces of patients with acute gastroenteritis. The norovirus genome is approximately 7.7 kb in length and contains three open reading frames (ORFs). ORF1 encodes non-structural proteins, including RNA-dependent RNA polymerase (RdRp); ORF2 and ORF3 encode the major (VP1) and minor (VP2) capsid proteins, respectively. Based on the phylogenetic analysis of the gene sequence, norovirus can be divided into six genogroups (GI-GVI), each of which is further divided into multiple genotypes. Human infections are mostly caused by GI and GII genogroups, with GII being the predominant genotype; statistically, over 75% of human Norovirus acute gastroenteritis cases are caused by GII infection.

[0004] The detection rate and prevalence of different genotypes vary considerably across different periods and regions. Over the past 20 years, the GII.4 genotype has been the dominant prevalence in outbreaks and sporadic cases worldwide. Other genotypes, such as GII.2, GII.3, GII.6, and GII.17, have higher detection rates than GII.4 and have even caused small-scale outbreaks in some local areas in recent years due to changes in the epidemic trend.

[0005] The diverse and rapidly mutating genotypes of norovirus, coupled with the lack of ideal cell culture systems and animal models, have significantly limited the development of antiviral drugs. To date, effective drugs, vaccines, and preventative measures remain scarce. Therefore, developing effective antiviral drugs and preventative vaccines is a pressing issue that urgently needs to be addressed. Summary of the Invention

[0006] Therefore, the purpose of this invention is to address the shortcomings of existing technologies by providing a virus-like particle (VLP) that targets two valence types of norovirus. The inventors have conducted extensive research on cross-immunity between antigens of different norovirus valence types and discovered that GII.6 norovirus VLPs also exhibit cross-immunity against GII.7. Based on this discovery, the inventors provide a GII.6 norovirus VLP and its uses.

[0007] The objective of this invention is achieved through the following technical solution:

[0008] On one hand, the present invention provides a norovirus virus-like particle or an active fragment thereof, said virus-like particle or active fragment comprising or consisting of an amino acid sequence selected from or composed of:

[0009] (1) The amino acid sequence shown in SEQ ID NO:5;

[0010] (2) An amino acid sequence having at least 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99% or higher identity with the amino acid sequence shown in SEQ ID NO:5;

[0011] (3) An amino acid sequence having one or more amino acid substitutions, deletions, or insertions at one or more sites in the amino acid sequence shown in SEQ ID NO:5; and

[0012] (4) The amino acid sequence shown in SEQ ID NO:5 consists of the Xth to the 547th consecutive amino acid, where X is an integer between 2 and 27; preferably, X is 27.

[0013] On the other hand, the present invention provides a composition for inducing an immune response to norovirus in animals, comprising norovirus virus-like particles or active fragments thereof as described in the present invention.

[0014] In the composition according to the present invention, the animal may be a mammal, preferably a human.

[0015] According to the composition of the present invention, the norovirus is GII.6 and / or GII.7 norovirus.

[0016] Preferably, the composition according to the present invention is a pharmaceutical composition for the prevention and / or treatment of norovirus infection, such as a vaccine.

[0017] Preferably, the pharmaceutical composition further comprises one or more adjuvants selected from aluminum adjuvants, TRL adjuvants, and saponin adjuvants. More preferably, the aluminum adjuvant is selected from one or more of aluminum hydroxide, aluminum phosphate, and aluminum sulfate; even more preferably, the aluminum adjuvant is aluminum hydroxide; more preferably, the TRL adjuvant is TRL9 adjuvant; even more preferably, the TRL9 adjuvant is CPG adjuvant; even more preferably, the nucleotide sequence of the CPG adjuvant is selected from one of SEQ ID NO:9, SEQ ID NO:10, or SEQ ID NO:11; more preferably, the saponin adjuvant is QS21 or Iscom adjuvant.

[0018] According to the composition of the present invention, the norovirus infection is gastroenteritis, preferably viral acute gastroenteritis.

[0019] Furthermore, the present invention also provides an immunoassay kit comprising norovirus virus-like particles or active fragments thereof as described in the present invention, or a composition as described in the present invention. Preferably, the kit is a kit for detecting norovirus.

[0020] Furthermore, the present invention also provides the use of norovirus virus-like particles or active fragments thereof as described in the present invention, the compositions described in the present invention, or the kits described in the present invention in the preparation of the following products:

[0021] (1) Medications used to prevent and / or treat norovirus infection;

[0022] (2) A kit for diagnosing norovirus infection; or

[0023] (3) Immunogen composition for developing norovirus antibodies.

[0024] According to the use described in this invention, the norovirus infection is gastroenteritis, preferably viral acute gastroenteritis.

[0025] The inventors of this invention have discovered that GII.6 norovirus virus-like particles containing the amino acid sequence described in this invention also induce cross-immunity against GII.7 norovirus, with a stronger immune response. This achieves the goal of using a single antigen to prevent, treat, and / or diagnose two valence types of norovirus infection, significantly reducing the amount of antigen required. Using the single antigen of this invention to cover two valence types reduces the complexity of pharmaceutical composition or vaccine preparation processes, lowers production costs, and has broad market prospects. Attached Figure Description

[0026] The embodiments of the present invention will now be described in detail with reference to the accompanying drawings, wherein:

[0027] Figure 1This study demonstrated the cross-immunity of GII.6 norovirus virus-like particles (VLPs) against norovirus of various valences.

[0028] Figure 2 The immunogenicity of virus-like particles (VLPs) of GII.6 norovirus against the said valence norovirus type with cross-immunity was demonstrated.

[0029] definition:

[0030] Unless otherwise defined, all technical terms used herein have the same meaning as understood by one of ordinary skill in the art. For definitions of terms used in this field, those skilled in the art may refer to Current Protocols in Molecular Biology (Ausubel). The abbreviations for amino acid residues are the standard 3-letter and / or 1-letter codes used in this field to refer to one of the 20 commonly used L-amino acids.

[0031] It should also be noted that, as used in this specification, the term "or" may be used interchangeably with the term "and / or" unless the context clearly indicates otherwise.

[0032] As used herein, the terms "pharmaceutical composition," "combination drug," and "pharmaceutical combination" are used interchangeably to refer to a combination of at least one drug substance and optional pharmaceutically acceptable excipients or adjuvants combined together to achieve a particular purpose. In some embodiments, the pharmaceutical composition comprises combinations that are separate in time and / or space, provided that they can work together to achieve the objectives of the invention. For example, the components contained in the pharmaceutical composition (e.g., GII.6 or Al(OH)3) may be applied to the subject as a whole or separately. When the components contained in the pharmaceutical composition are applied to the subject separately, the components may be applied to the subject simultaneously or sequentially.

[0033] As used herein, the term "CpG oligodeoxynucleotide" or "CpG-ODN" refers to a short, single-stranded synthetic DNA molecule containing one or more "CpG" units, where C represents cytosine, G represents guanine, and p represents a phosphodiester bond. Specifically, the CpG oligodeoxynucleotide is unmethylated. In some embodiments, the CpG-ODN comprises phosphate thioester links or a phosphate thioester backbone. That is, in some embodiments, the CpG-ODN is a phosphate thioester oligodeoxynucleotide (i.e., a thioolipodeoxynucleotide). Preferably, all nucleotide linkages in the CpG-ODN are phosphate thioester links, i.e., the CpG-ODN is a fully thioolipodeoxynucleotide. In other embodiments, the CpG-ODN contains two or more copies of the 5'-TTCGTT-3' motif or the 5'-TCGTCGTCG-3' motif. Specifically, the CpG-ODN has a sequence selected from the following: TCG TTC GTT CGT TCG TTC GTT (SEQ ID NO:9), TCG TTC GTTCGT TCG TTC GTT CGT T (SEQ ID NO:10) or TCG TCG TCG TCG TCG TCG TCG (SEQ ID NO:11), preferably TCG TTC GTT CGT TCG TTC GTT (SEQ ID NO:9).

[0034] The term "saponin" as used in this article refers to the active ingredient found in the corresponding plant, especially QS21, which is a saponin from a soap tree, or Iscom adjuvant, which is an immunostimulatory complex adjuvant. Specifically, it refers to Iscom matrix (ISCOMMATRIX) without antigen, which is an adjuvant with a cage-like structure composed of phospholipids, saponins, and cholesterol.

[0035] As used in this article, "therapeutic effective dose" or "effective dose" refers to a dose sufficient to demonstrate its benefit to the recipient. The actual amount administered, as well as the rate and duration of administration, depends on the individual's condition and severity. Prescribing treatment (e.g., determining the dosage) is ultimately the responsibility of and depends on the general practitioner and other physicians, who typically consider the disease being treated, the individual patient's condition, the site of delivery, the method of administration, and other factors known to the physician.

[0036] The term “mammal” as used in this article refers to humans, but can also refer to other animals, such as wild animals (e.g., herons, storks, cranes, etc.), livestock (e.g., ducks, geese, etc.) or laboratory animals (e.g., orangutans, monkeys, rats, mice, rabbits, guinea pigs, marmots, ground squirrels, etc.).

[0037] In other embodiments, the compositions of the present invention may also contain additional additives, such as pharmaceutically acceptable carriers or additives, especially when they are in the form of pharmaceutical formulations.

[0038] Preferred drug carriers are, in particular, water, buffered aqueous solutions, and preferably isotonic salt solutions such as PBS (phosphate-buffered saline), glucose, mannitol, dextran, lactose, starch, magnesium stearate, cellulose, magnesium carbonate, 0.3% glycerol, hyaluronic acid, ethanol, or polyalkylene glycols such as polypropylene glycol, triglycerides, etc. The type of drug carrier used depends particularly on whether the composition according to the invention is formulated for oral, nasal, intradermal, subcutaneous, intramuscular, or intravenous administration. The compositions according to the invention may contain wetting agents, emulsifiers, or buffering substances as additives.

[0039] The pharmaceutical compositions, vaccines, or pharmaceutical preparations according to the present invention may be administered by any suitable route, such as oral, nasal, intradermal, subcutaneous, intramuscular, or intravenous administration. Detailed Implementation

[0040] The present invention will be further described below with reference to the accompanying drawings and through a description of specific embodiments. However, this is not a limitation of the present invention. Those skilled in the art can make various modifications or improvements based on the basic idea of ​​the present invention, but as long as they do not depart from the basic idea of ​​the present invention, they are all within the scope of the present invention.

[0041] Example 1: Preparation method of norovirus antigen

[0042] Multiple valence norovirus VLP proteins were prepared, and their amino acid sequences are shown below:

[0043] The amino acid sequence of the GII.1 type norovirus VLP protein is shown in SEQ ID NO:1.

[0044] The amino acid sequence of the GII.2 type norovirus VLP protein is shown in SEQ ID NO:2.

[0045] The amino acid sequence of the GII.3 type norovirus VLP protein is shown in SEQ ID NO:3.

[0046] The amino acid sequence of the GII.4 type norovirus VLP protein is shown in SEQ ID NO:4.

[0047] The amino acid sequence of the GII.6 norovirus VLP protein is shown in SEQ ID NO:5.

[0048] The amino acid sequence of the GII.7 type norovirus VLP protein is shown in SEQ ID NO:6.

[0049] The amino acid sequence of the GII.12 norovirus VLP protein is shown in SEQ ID NO:7.

[0050] The amino acid sequence of the GII.17 norovirus VLP protein is shown in SEQ ID NO:8.

[0051] The specific preparation method is as follows: The nucleic acid sequence of the target protein is optimized according to the target protein sequence to make its codons conform to the yeast expression system, and the target gene is synthesized; the synthesized target gene is ligated to the pMAUR(SC)KARS1 plasmid by enzyme digestion and ligation, transformed into Top 10 competent cells, positive single clones are selected, and the positive single clones are sequenced for verification; the single clone cells are amplified in large quantities, and plasmids that are suitable for electroporation are extracted in large quantities using a plasmid extraction kit; the plasmid is transformed into uracil auxotrophic Hansenula polymorpha competent cells by electroporation, and positive transformed cells are obtained by G418 resistance selection.

[0052] The recombinant strain was activated by inoculating yeast extract peptone glucose medium. The activated strain was then transferred to yeast extract peptone glucose medium for further culture. After the strain reached the plateau phase, methanol was added to the medium for induction culture twice, every 24 hours. After induction, the bacterial cells were collected, lysed, and purified to obtain the protein.

[0053] Example 2 Animal Immunization Experiment

[0054] 2.1 Laboratory Animals

[0055] Balb / c mice, female, 6 weeks old, 12 mice, Shanghai Lingchang Laboratory Animal Technology Co., Ltd.

[0056] 2.2 Experimental Materials

[0057] The VLP proteins of various valences of norovirus were prepared in Example 1.

[0058] Aluminum hydroxide: purchased from Beijing Bio-Lab Technology Co., Ltd.

[0059] 2.3 Experimental Grouping

[0060] Table 1 Grouping of Animal Immunization Experiments

[0061]

[0062] 2.4 Animal Immunization

[0063] 1) Route of administration: Intramuscular injection.

[0064] 2) Administration method: Select the left hind thigh of a mouse and inject 100 μl / mouse.

[0065] 3) Dosing frequency and duration: once every 3 weeks, for a total of 3 doses, i.e., injections are given in week 0, week 3 and week 6 respectively. Blood samples are collected two weeks after the first immunization, three weeks after the second immunization, and two weeks after the third immunization as the final endpoint.

[0066] 4) Serum collection method: Place whole blood in a 37℃ constant temperature incubator for 40-50 min, then incubate at 4℃ for 1 h, centrifuge at 12000 rpm at 4℃ for 10 min, collect the supernatant, and freeze it in a medical low temperature storage box for later use.

[0067] Example 3 Cross-blocking experiment of the drug composition

[0068] 3.1 Reagents and Materials

[0069] 1) BSA (bovine serum albumin, Beyotime), TMB chromogenic solution (Thermo Fisher Scientific), HBGA (Glycotech), rabbit polyantiserum (Jingtiancheng Biotechnology), goat anti-rabbit IgG-HRP (horseradish peroxidase-labeled goat anti-rabbit IgG, SIGMA), avidin plate (Thermo Fisher Scientific).

[0070] 2) Washing buffer: 0.1M PB buffer

[0071] Diluent: 0.1M PB buffer + 0.25% BSA

[0072] Stop solution: 2M H2SO4

[0073] All were prepared by Nanjing Yuandaweixin Biotechnology Co., Ltd.

[0074] 3) Norovirus VLP proteins: G1, GII.2, GII.3, GII.4, GII.6, GII.7, GII.12 and GII.17, prepared in Example 1, 8 μg / ml

[0075] 3.2 Testing Procedures

[0076] 1) Wash the affinity plate once with 200 μl / well washing solution;

[0077] 2) Dilute HBGA to 4 μg / ml and add 100 μl / well to the avidin plate from step 1), and incubate at 25°C for 1 h;

[0078] 3) In a 96-well plate, the serum was serially diluted from 20-fold to 1280-fold, with a final volume of 60 μl. 60 μl of monovalent norovirus VLP protein was added to each well to form a VLP-serum mixture. A positive control (no serum) and a blank control (diluent) were set up. The mixture was incubated at 4°C for 1 h.

[0079] 4) Wash the avidin plate coated with HBGA in step 2) twice with 200 μl / well washing buffer. Take 100 μl of VLP-serum mixture from the 96-well plate in step 3) and add it to the avidin plate. Incubate at 4°C for 2 h.

[0080] 5) Wash the avidin plate three times with 200 μl / well washing buffer, add rabbit antiserum (1:10000), and incubate at 4°C for 1 h;

[0081] 6) Wash the avidin plate three times with 200 μl / well washing buffer, add goat anti-rabbit IgG-HRP (1:10000), and incubate at 37°C for 40 min;

[0082] 7) Wash the affinity plate three times with 200 μl / well washing buffer, add 100 μl / well TMB colorimetric solution, and develop the color at 25℃ for 5-15 min;

[0083] 8) Termination: Add 100 μl of stop solution per well to terminate the reaction;

[0084] 9) Reading: Measure the OD450nm value (corrected to OD630nm).

[0085] Detection standard: Blocking index % = (1 - serum group A value / positive control A value) × 100%, calculate BT50, which is the highest serum dilution that can block 50% VLP from binding to HBGA.

[0086] Set a cut-off value: Use twice the BT50 of Al(OH)3 group as the cut-off value to determine whether there is cross-immunization.

[0087] 3.3 Experimental Results

[0088] Experimental results are as follows Figure 1 As shown: The GII.6 provided by this invention has significant cross-immunity against GII.7, and the BT50 can reach about 60.

[0089] Example 4: Detection of IgG antibody levels in the pharmaceutical composition

[0090] 4.1 Reagents and Materials

[0091] 1) Skim milk powder (BD Pharmaceuticals), PBS (Zhongshan Jinqiao Pharmaceuticals), carbonate (SIGMA Pharmaceuticals), Tween 20 (BIO-LINK Pharmaceuticals), TMB chromogenic solution (Thermo Pharmaceuticals), horseradish peroxidase-labeled goat anti-rabbit IgG (hereinafter referred to as goat anti-rabbit IgG-HRP, SIGMA Pharmaceuticals)

[0092] 2) Coating solution: 0.5% (g / 100ml) carbonate solution

[0093] Washing buffer PBST: 10 g / L PBS + 0.5 ml / L Tween 20

[0094] Diluent: 2% skim milk PBST

[0095] Stop solution: 2M H2SO4

[0096] All of the above were prepared by Nanjing Yuansai Weixin Biomedical Co., Ltd.

[0097] 4.2 Testing Procedures

[0098] 1) Coating: Norovirus VLP proteins of types GI.1, GII.2, GII.3, GII.4, GII.6, GII.7, GII.12 and GII.17 were diluted to 1 μg / ml with coating buffer and coated on ELISA plates, 50 μl / well, overnight at 4°C;

[0099] 2) Washing: Wash the plate 3 times with PBST washing buffer, 2-3 min each time;

[0100] 3) Blocking: Add 5% milk blocking solution, 200 μl / well, and block at 37℃ for 1 h;

[0101] 4) Washing: Wash the plate 3 times with PBST washing buffer, 2-3 min each time;

[0102] 5) Primary antibody: Add serially diluted serum, starting from 90-fold and serially diluted 3-fold to 196830-fold, 50 μl / well, set up two blank control wells, and incubate at 37℃ for 1 h;

[0103] 6) Washing: Wash the plate 3 times with PBST washing buffer, 2-3 min each time;

[0104] 7) Secondary antibody: HRP-goat anti-mouse IgG antibody (1:10000), incubated at 37℃ for 40 min;

[0105] 8) Washing: Wash the plate 3 times with PBST washing buffer, 2-3 min each time;

[0106] 9) Color development: Add 50 μl of TMB color development solution to each well and develop for 10 min;

[0107] 10) Termination: 50 μl of stop solution per well to terminate the reaction;

[0108] 11) Reading: OD450nm value measured by enzyme-linked immunosorbent assay (ELISA) (corrected to OD630nm).

[0109] 3.6 Experimental Results

[0110] The results are as follows Figure 2As shown: The GII.6 virus-like particle (VLP) provided by this invention exhibits an immune response against GII.7, and the antibody titer against GII.7 can reach a level of over 10,000.

[0111] In summary, this invention provides the use of virus-like particles (VLPs) of GII.6 norovirus for the preparation of drugs or kits for the prevention, treatment, and / or diagnosis of GII.7 norovirus infection. These virus-like particles exhibit strong cross-immunization against other valence antigens, and can be used to prepare vaccines covering multiple valences, achieving the goal of using a single antigen to cover multiple valences. This can reduce production costs and has broad market prospects.

[0112] Although the present invention has been described in detail above, those skilled in the art will understand that various modifications and changes can be made to the present invention without departing from its spirit and scope. The scope of the present invention is not limited to the detailed description above, and such modifications and changes should fall within the scope of the claims. While only examples of specific embodiments of the present invention have been described above, those skilled in the art should understand that these are merely illustrative examples, and the scope of protection of the present invention is defined by the appended claims. Those skilled in the art can make various changes or modifications to these embodiments without departing from the principles and essence of the present invention, but all such changes or modifications should fall within the scope of protection of the present invention. sequence list <110> Nanjing Yuanda Weixin Biomedical Co., Ltd. <120> Norovirus virus-like particles and their uses <130> DIC20110101 <160> 11 <170> SIPOSequenceListing 1.0 <210> 1 <211> 530 <212> PRT <213> Norovirus <400> 1 Met Met Met Ala Ser Lys Asp Ala Thr Ser Ser Val Asp Gly Ala Ser 1 5 10 15 Gly Ala Gly Gln Leu Val Pro Glu Val Asn Ala Ser Asp Pro Leu Ala 20 25 30 Met Asp Pro Val Ala Gly Ser Ser Thr Ala Val Ala Thr Ala Gly Gln 35 40 45 Val Asn Pro Ile Asp Pro Trp Ile Ile Asn Asn Phe Val Gln Ala Pro 50 55 60 Gln Gly Glu Phe Thr Ile Ser Pro Asn Asn Thr Pro Gly Asp Val Leu 65 70 75 80 Phe Asp Leu Ser Leu Gly Pro His Leu Asn Pro Phe Leu Leu His Leu 85 90 95 Ser Gln Met Tyr Asn Gly Trp Val Gly Asn Met Arg Val Arg Ile Met 100 105 110 Leu Ala Gly Asn Ala Phe Thr Ala Gly Lys Ile Ile Val Ser Cys Ile 115 120 125 Pro Pro Gly Phe Gly Ser His Asn Leu Thr Ile Ala Gln Ala Thr Leu 130 135 140 Phe Pro His Val Ile Ala Asp Val Arg Thr Leu Asp Pro Ile Glu Val 145 150 155 160 Pro Leu Glu Asp Val Arg Asn Val Leu Phe His Asn Asn Asp Arg Asn 165 170 175 Gln Gln Thr Met Arg Leu Val Cys Met Leu Tyr Thr Pro Leu Arg Thr 180 185 190 Gly Gly Gly Thr Gly Asp Ser Phe Val Val Ala Gly Arg Val Met Thr 195 200 205 Cys Pro Ser Pro Asp Phe Asn Phe Leu Phe Leu Val Pro Pro Thr Val 210 215 220 Glu Gln Lys Thr Arg Pro Phe Thr Leu Pro Asn Leu Pro Leu Ser Ser 225 230 235 240 Leu Ser Asn Ser Arg Ala Pro Leu Pro Ile Ser Ser Met Gly Ile Ser 245 250 255 Pro Asp Asn Val Gln Ser Val Gln Phe Gln Asn Gly Arg Cys Thr Leu 260 265 270 Asp Gly Arg Leu Val Gly Thr Thr Pro Val Ser Leu Ser His Val Ala 275 280 285 Lys Ile Arg Gly Thr Ser Asn Gly Thr Val Ile Asn Leu Thr Glu Leu 290 295 300 Asp Gly Thr Pro Phe His Pro Phe Glu Gly Pro Ala Pro Ile Gly Phe 305 310 315 320 Pro Asp Leu Gly Gly Cys Asp Trp His Ile Asn Met Thr Gln Phe Gly 325 330 335 His Ser Ser Gln Thr Gln Tyr Asp Val Asp Thr Thr Pro Asp Thr Phe 340 345 350 Val Pro His Leu Gly Ser Ile Gln Ala Asn Gly Ile Gly Ser Gly Asn 355 360 365 Tyr Val Gly Val Leu Ser Trp Ile Ser Pro Pro Ser His Pro Ser Gly 370 375 380 Ser Gln Val Asp Leu Trp Lys Ile Pro Asn Tyr Gly Ser Ser Ile Thr 385 390 395 400 Glu Ala Thr His Leu Ala Pro Ser Val Tyr Pro Pro Gly Phe Gly Glu 405 410 415 Val Leu Val Phe Phe Met Ser Lys Met Pro Gly Pro Gly Ala Tyr Asn 420 425 430 Leu Pro Cys Leu Leu Pro Gln Glu Tyr Ile Ser His Leu Ala Ser Glu 435 440 445 Gln Ala Pro Thr Val Gly Glu Ala Ala Leu Leu His Tyr Val Asp Pro 450 455 460 Asp Thr Gly Arg Asn Leu Gly Glu Phe Lys Ala Tyr Pro Asp Gly Phe 465 470 475 480 Leu Thr Cys Val Pro Asn Gly Ala Ser Ser Gly Pro Gln Gln Leu Pro 485 490 495 Ile Asn Gly Val Phe Val Phe Val Ser Trp Val Ser Arg Phe Tyr Gln 500 505 510 Leu Lys Pro Val Gly Thr Ala Ser Ser Ala Arg Gly Arg Leu Gly Leu 515 520 525 Silver Silver 530 <210> 2 <211> 542 <212> PRT <213> Norovirus <400> 2 Met Lys Met Ala Ser Asn Asp Ala Ala Pro Ser Thr Asp Gly Ala Ala 1 5 10 15 Gly Leu Val Pro Glu Ser Asn Asn Glu Val Met Ala Leu Glu Pro Val 20 25 30 Ala Gly Ala Ala Leu Ala Ala Pro Val Thr Gly Gln Thr Asn Ile Ile 35 40 45 Asp Pro Trp Ile Arg Ala Asn Phe Val Gln Ala Pro Asn Gly Glu Phe 50 55 60 Thr Val Ser Pro Arg Asn Ala Pro Gly Glu Val Leu Leu Asn Leu Glu 65 70 75 80 Leu Gly Pro Glu Leu Asn Pro Tyr Leu Ala His Leu Ala Arg Met Tyr 85 90 95 Asn Gly Tyr Ala Gly Gly Met Glu Val Gln Val Met Leu Ala Gly Asn 100 105 110 Ala Phe Thr Ala Gly Lys Leu Val Phe Ala Ala Val Pro Pro His Phe 115 120 125 Pro Val Glu Asn Leu Ser Pro Gln Gln Ile Thr Met Phe Pro His Val 130 135 140 Ile Ile Asp Val Arg Thr Leu Glu Pro Val Leu Leu Pro Leu Pro Asp 145 150 155 160 Val Arg Asn Asn Phe Phe His Tyr Asn Gln Lys Asp Asp Pro Lys Met 165 170 175 Arg Ile Val Ala Met Leu Tyr Thr Pro Leu Arg Ser Asn Gly Ser Gly 180 185 190 Asp Asp Val Phe Thr Val Ser Cys Arg Val Leu Thr Arg Pro Ser Pro 195 200 205 Asp Phe Asp Phe Thr Tyr Leu Val Pro Pro Thr Val Glu Ser Lys Thr 210 215 220 Lys Pro Phe Thr Leu Pro Ile Leu Thr Leu Gly Glu Leu Ser Asn Ser 225 230 235 240 Arg Phe Pro Val Ser Ile Asp Gln Met Tyr Thr Ser Pro Asn Glu Ile 245 250 255 Ile Ser Val Gln Cys Gln Asn Gly Arg Cys Thr Leu Asp Gly Glu Leu 260 265 270 Gln Gly Thr Thr Gln Leu Gln Val Ser Gly Ile Cys Ala Phe Lys Gly 275 280 285 Glu Val Thr Ala His Leu His Asp Asn Asp His Leu Tyr Asn Val Thr 290 295 300 Ile Thr Asn Leu Asn Gly Ser Pro Phe Asp Pro Ser Glu Asp Ile Pro 305 310 315 320 Ala Pro Leu Gly Val Pro Asp Phe Gln Gly Arg Val Phe Gly Ile Ile 325 330 335 Ser Gln Arg Asp Lys His Asn Ser Pro Gly His Asn Glu Pro Ala Asn 340 345 350 Arg Gly His Asp Ala Val Val Pro Thr Tyr Thr Ala Gln Tyr Thr Pro 355 360 365 Lys Leu Gly Gln Ile Gln Ile Gly Thr Trp Gln Thr Asp Asp Leu Thr 370 375 380 Val Asn Gln Pro Val Lys Phe Thr Pro Val Gly Leu Asn Asp Thr Glu 385 390 395 400 His Phe Asn Gln Trp Val Val Pro Arg Tyr Ala Gly Ala Leu Asn Leu 405 410 415 Asn Thr Asn Leu Ala Pro Ser Val Ala Pro Val Phe Pro Gly Glu Arg 420 425 430 Leu Leu Phe Phe Arg Ser Tyr Ile Pro Leu Lys Gly Gly Tyr Gly Asn 435 440 445 Pro Ala Ile Asp Cys Leu Leu Pro Gln Glu Trp Val Gln His Phe Tyr 450 455 460 Gln Glu Ala Ala Pro Ser Met Ser Glu Val Ala Leu Val Arg Tyr Ile 465 470 475 480 Asn Pro Asp Thr Gly Arg Ala Leu Phe Glu Ala Lys Leu His Arg Ala 485 490 495 Gly Phe Met Thr Val Ser Ser Asn Thr Ser Ala Pro Val Val Val Pro 500 505 510 Ala Asn Gly Tyr Phe Arg Phe Asp Ser Trp Val Asn Gln Phe Tyr Ser 515 520 525 Leu Ala Pro Met Gly Thr Gly Asn Gly Arg Arg Arg Val Gln 530 535 540 <210> 3 <211> 548 <212> PRT <213> Norovirus <400> 3 Met Lys Met Ala Ser Asn Asp Ala Thr Pro Ser Asn Asp Gly Ala Ala 1 5 10 15 Gly Leu Val Pro Glu Ile Asn Asn Glu Ala Met Ala Leu Asp Pro Val 20 25 30 Ala Gly Ala Ala Ile Ala Ala Pro Leu Thr Gly Gln Gln Asn Ile Ile 35 40 45 Asp Pro Trp Ile Met Asn Asn Phe Val Gln Ala Pro Gly Gly Glu Phe 50 55 60 Thr Val Ser Pro Arg Asn Ser Pro Gly Glu Val Leu Leu Asn Leu Glu 65 70 75 80 Leu Gly Pro Glu Ile Asn Pro Tyr Leu Ala His Leu Ala Arg Met Tyr 85 90 95 Asn Gly Tyr Ala Gly Gly Phe Glu Val Gln Val Val Leu Ala Gly Asn 100 105 110 Ala Phe Thr Ala Gly Lys Ile Ile Phe Ala Ala Ile Pro Pro Asn Phe 115 120 125 Pro Ile Asp Asn Leu Ser Ala Ala Gln Ile Thr Met Cys Pro His Val 130 135 140 Ile Val Asp Val Arg Gln Leu Glu Pro Val Asn Leu Pro Met Pro Asp 145 150 155 160 Val Arg Asn Asn Phe Phe His Tyr Asn Gln Gly Ser Asp Ser Arg Leu 165 170 175 Arg Leu Val Ala Met Leu Tyr Thr Pro Leu Arg Ala Asn Asn Ser Gly 180 185 190 Asp Asp Val Phe Thr Val Ser Cys Arg Val Leu Thr Arg Pro Ser Pro 195 200 205 Glu Phe Ser Phe Asn Phe Leu Val Pro Pro Thr Val Glu Ser Lys Thr 210 215 220 Lys Pro Phe Thr Leu Pro Ile Leu Thr Ile Ser Glu Met Ser Asn Ser 225 230 235 240 Arg Phe Pro Val Pro Ile Asp Ser Leu His Thr Ser Pro Thr Glu Asn 245 250 255 Ile Val Val Gln Cys Gln Asn Gly Arg Val Thr Leu Asp Gly Glu Leu 260 265 270 Met Gly Thr Thr Gln Leu Leu Pro Ser Gln Ile Cys Ala Phe Arg Gly 275 280 285 Val Leu Thr Arg Ser Thr Ser Arg Ala Ser Asp Gln Ala Asp Thr Ala 290 295 300 Thr Pro Arg Leu Phe Asn Tyr Tyr Trp His Ile Gln Leu Asp Asn Leu 305 310 315 320 Asn Gly Thr Pro Tyr Asp Pro Ala Glu Asp Ile Pro Gly Pro Leu Gly 325 330 335 Thr Pro Asp Phe Arg Gly Lys Val Phe Gly Val Ala Ser Gln Arg Asn 340 345 350 Pro Asp Ser Thr Thr Arg Ala His Glu Ala Lys Val Asp Thr Thr Ala 355 360 365 Gly Arg Phe Thr Pro Lys Leu Gly Ser Leu Glu Ile Ser Thr Glu Ser 370 375 380 Gly Asp Phe Asp Gln Asn Gln Pro Thr Arg Phe Thr Pro Val Gly Ile 385 390 395 400 Gly Val Asp His Glu Ser Asp Phe Gln Gln Trp Ser Leu Pro Asp Tyr 405 410 415 Ser Gly Gln Phe Thr His Asn Met Asn Leu Ala Pro Ala Val Ala Pro 420 425 430 Asn Phe Pro Gly Glu Gln Leu Leu Phe Phe Arg Ser Gln Leu Pro Ser 435 440 445 Ser Gly Gly Arg Ser Asn Gly Ile Leu Asp Cys Leu Val Pro Gln Glu 450 455 460 Trp Val Gln His Phe Tyr Gln Glu Ser Ala Pro Ala Gln Thr Gln Val 465 470 475 480 Ala Leu Val Arg Tyr Val Asn Pro Asp Thr Gly Arg Val Leu Phe Glu 485 490 495 Ala Lys Leu His Lys Leu Gly Phe Met Thr Ile Ala Lys Asn Gly Asp 500 505 510 Ser Pro Ile Thr Val Pro Pro Asn Gly Tyr Phe Arg Phe Glu Ser Trp 515 520 525 Val Asn Pro Phe Tyr Thr Leu Ala Pro Met Gly Thr Gly Asn Gly Arg 530 535 540 Arg Arg Val Gln 545 <210> 4 <211> 540 <212> PRT <213> Norovirus <400> 4 Met Lys Met Ala Ser Ser Asp Ala Asn Pro Ser Asp Gly Ser Ala Ala 1 5 10 15 Asn Leu Val Pro Glu Val Asn Asn Glu Val Met Ala Leu Glu Pro Val 20 25 30 Val Gly Ala Ala Ile Ala Ala Pro Val Ala Gly Gln Gln Asn Val Ile 35 40 45 Asp Pro Trp Ile Arg Asn Asn Phe Val Gln Ala Pro Gly Gly Glu Phe 50 55 60 Thr Val Ser Pro Arg Asn Ala Pro Gly Glu Ile Leu Trp Ser Ala Pro 65 70 75 80 Leu Gly Pro Asp Leu Asn Pro Tyr Leu Ser His Leu Ala Arg Met Tyr 85 90 95 Asn Gly Tyr Ala Gly Gly Phe Glu Val Gln Val Ile Leu Ala Gly Asn 100 105 110 Ala Phe Thr Ala Gly Lys Val Ile Phe Ala Ala Val Pro Pro Asn Phe 115 120 125 Pro Thr Glu Gly Leu Ser Pro Ser Gln Val Thr Met Phe Pro His Ile 130 135 140 Val Val Asp Val Arg Gln Leu Glu Pro Val Leu Ile Pro Leu Pro Asp 145 150 155 160 Val Arg Asn Asn Phe Tyr His Tyr Asn Gln Ser Asn Asp Pro Thr Ile 165 170 175 Lys Leu Ile Ala Met Leu Tyr Thr Pro Leu Arg Ala Asn Asn Ala Gly 180 185 190 Asp Asp Val Phe Thr Val Ser Cys Arg Val Leu Thr Arg Pro Ser Pro 195 200 205 Asp Phe Asp Phe Ile Phe Leu Val Pro Pro Thr Val Glu Ser Arg Thr 210 215 220 Lys Pro Phe Ser Val Pro Val Leu Thr Val Glu Glu Met Thr Asn Ser 225 230 235 240 Arg Phe Pro Ile Pro Leu Glu Lys Leu Phe Thr Gly Pro Ser Ser Ala 245 250 255 Phe Val Val Gln Pro Gln Asn Gly Arg Cys Thr Thr Asp Gly Val Leu 260 265 270 Leu Gly Thr Thr Gln Leu Ser Pro Val Asn Ile Cys Thr Phe Arg Gly 275 280 285 Asp Val Thr His Ile Thr Gly Ser Arg Asn Tyr Thr Met Asn Leu Ala 290 295 300 Ser Gln Asn Trp Asn Asn Tyr Asp Pro Thr Glu Glu Ile Pro Ala Pro 305 310 315 320 Leu Gly Thr Pro Asp Phe Val Gly Lys Ile Gln Gly Val Leu Thr Gln 325 330 335 Thr Thr Arg Thr Asp Gly Ser Thr Arg Gly His Lys Ala Thr Val Tyr 340 345 350 Thr Gly Ser Ala Asp Phe Ala Pro Lys Leu Gly Arg Val Gln Phe Glu 355 360 365 Thr Asp Thr Asp His Asp Phe Glu Ala Asn Gln Asn Thr Lys Phe Thr 370 375 380 Pro Val Gly Val Ile Gln Asp Gly Ser Thr Thr His Arg Asn Glu Pro 385 390 395 400 Gln Gln Trp Val Leu Pro Ser Tyr Ser Gly Arg Asn Thr His Asn Val 405 410 415 His Leu Ala Pro Ala Val Ala Pro Thr Phe Pro Gly Glu Gln Leu Leu 420 425 430 Phe Phe Arg Ser Thr Met Pro Gly Cys Ser Gly Tyr Pro Asn Met Asp 435 440 445 Leu Asp Cys Leu Leu Pro Gln Glu Trp Val Gln Tyr Phe Tyr Gln Glu 450 455 460 Ala Ala Pro Ala Gln Ser Asp Val Ala Leu Leu Arg Phe Val Asn Pro 465 470 475 480 Asp Thr Gly Arg Val Leu Phe Glu Cys Lys Leu His Lys Ser Gly Tyr 485 490 495 Val Thr Val Ala His Thr Gly Gln His Asp Leu Val Ile Pro Pro Asn 500 505 510 Gly Tyr Phe Arg Phe Asp Ser Trp Val Asn Gln Phe Tyr Thr Leu Ala 515 520 525 Pro Met Gly Asn Gly Thr Gly Arg Arg Arg Ala Val 530 535 540 <210> 5 <211> 547 <212> PRT <213> Norovirus <400> 5 Met Lys Met Ala Ser Asn Asp Ala Ala Pro Ser Asn Asp Gly Ala Ala 1 5 10 15 Asn Leu Val Pro Glu Ala Thr Asn Glu Val Met Ala Leu Glu Pro Val 20 25 30 Val Gly Ala Ser Ile Ala Ala Pro Val Val Gly Gln Gln Asn Ile Ile 35 40 45 Asp Pro Trp Ile Arg Glu Asn Phe Val Gln Ala Pro Gln Gly Glu Phe 50 55 60 Thr Val Ser Pro Arg Asn Ser Pro Gly Glu Met Leu Leu Asn Leu Glu 65 70 75 80 Leu Gly Pro Glu Leu Asn Pro Tyr Leu Ser His Leu Ser Arg Met Tyr 85 90 95 Asn Gly Tyr Ala Gly Gly Met Gln Val Gln Val Val Leu Ala Gly Asn 100 105 110 Ala Phe Thr Ala Gly Lys Ile Ile Phe Ala Ala Val Pro Pro His Phe 115 120 125 Pro Val Glu Asn Ile Ser Ala Ala Gln Ile Thr Met Cys Pro His Val 130 135 140 Ile Val Asp Val Arg Gln Leu Glu Pro Val Leu Leu Pro Leu Pro Asp 145 150 155 160 Ile Arg Asn Arg Phe Phe His Tyr Asn Gln Glu Asn Thr Pro Arg Met 165 170 175 Arg Leu Val Ala Met Leu Tyr Thr Pro Leu Arg Ala Asn Ser Gly Glu 180 185 190 Asp Val Phe Thr Val Ser Cys Arg Val Leu Thr Arg Pro Ala Pro Asp 195 200 205 Phe Glu Phe Thr Phe Leu Val Pro Pro Thr Val Glu Ser Lys Thr Lys 210 215 220 Pro Phe Thr Leu Pro Ile Leu Thr Leu Gly Glu Leu Ser Asn Ser Arg 225 230 235 240 Phe Pro Ala Pro Ile Asp Met Leu Tyr Thr Asp Pro Asn Glu Ala Ile 245 250 255 Val Val Gln Pro Gln Asn Gly Arg Cys Thr Leu Asp Gly Thr Leu Gln 260 265 270 Gly Thr Thr Gln Leu Val Pro Thr Gln Ile Cys Ser Phe Arg Gly Thr 275 280 285 Leu Val Ser Gln Thr Ser Arg Ser Ala Asp Ser Thr Asp Ser Ala Pro 290 295 300 Arg Val Arg Ser His Pro Leu His Val Gln Leu Lys Asn Leu Asp Gly 305 310 315 320 Thr Pro Tyr Asp Pro Thr Asp Glu Val Pro Ala Val Leu Gly Ala Ile 325 330 335 Asp Phe Lys Gly Thr Val Phe Gly Val Ala Ser Gln Arg Asn Thr Thr 340 345 350 Gly Asn Ser Ile Gly Ala Thr Arg Ala His Glu Val His Ile Asp Thr 355 360 365 Thr Asn Pro Arg Tyr Thr Pro Lys Leu Gly Ser Val Leu Met Tyr Ser 370 375 380 Glu Ser Asn Asp Phe Asp Asp Gly Gln Pro Thr Arg Phe Thr Pro Ile 385 390 395 400 Gly Met Gly Ala Asp Asp Trp His Gln Trp Glu Leu Pro Glu Tyr Ser 405 410 415 Gly His Leu Thr Leu Asn Met Asn Leu Ala Pro Ala Leu Ala Pro Ala 420 425 430 Phe Pro Gly Glu Arg Ile Leu Phe Phe Arg Ser Val Val Pro Ser Ala 435 440 445 Gly Gly Tyr Gly Ser Gly His Ile Asp Cys Leu Ile Pro Gln Glu Trp 450 455 460 Val Gln His Phe Tyr Gln Glu Ala Ala Pro Ser Gln Ser Ala Val Ala 465 470 475 480 Leu Ile Arg Tyr Val Asn Pro Asp Thr Gly Arg Asn Ile Phe Glu Ala 485 490 495 Lys Leu His Arg Glu Gly Phe Ile Thr Val Ala Asn Ser Gly Asn Asn 500 505 510 Pro Ile Val Val Pro Pro Asn Gly Tyr Phe Arg Phe Glu Ala Trp Val 515 520 525 Asn Gln Phe Tyr Thr Leu Thr Pro Met Gly Thr Gly Gln Gly Arg Arg 530 535 540 Arg Val Gln 545 <210> 6 <211> 540 <212> PRT <213> Norovirus <400> 6 Met Lys Met Ala Ser Asn Asp Ala Ala Pro Ser Asn Asp Gly Ala Ala 1 5 10 15 Gly Leu Val Pro Glu Ile Asn Asn Glu Val Met Pro Leu Glu Pro Val 20 25 30 Ala Gly Ala Ser Leu Ala Thr Pro Val Ala Gly Gln His Asn Ile Ile 35 40 45 Asp Pro Trp Ile Arg Asn Asn Phe Val Gln Ala Pro Ala Gly Glu Phe 50 55 60 Thr Val Ser Pro Arg Asn Ser Pro Gly Glu Ile Leu Leu Asp Leu Glu 65 70 75 80 Leu Gly Pro Glu Leu Asn Pro Tyr Leu Ala His Leu Ala Arg Met Tyr 85 90 95 Asn Gly His Ala Gly Gly Met Glu Val Gln Ile Val Leu Ala Gly Asn 100 105 110 Ala Phe Thr Ala Gly Lys Ile Ile Phe Ala Ala Ile Pro Pro Gly Phe 115 120 125 Pro Tyr Glu Asn Leu Ser Pro Ser Gln Ile Thr Met Cys Pro His Val 130 135 140 Ile Ile Asp Val Arg Gln Leu Glu Pro Val Leu Leu Pro Met Pro Asp 145 150 155 160 Ile Arg Asn Asn Phe Phe His Tyr Asn Gln Ser Asn Asp Pro Lys Leu 165 170 175 Arg Leu Leu Ala Met Leu Tyr Thr Pro Leu Arg Ala Asn Asn Ser Gly 180 185 190 Asp Asp Val Phe Thr Val Ser Cys Arg Val Leu Thr Lys Pro Ser Pro 195 200 205 Asp Phe Glu Phe Thr Phe Leu Val Pro Pro Thr Val Glu Ser Lys Thr 210 215 220 Lys Gln Phe Thr Leu Pro Ile Leu Lys Ile Ser Glu Met Thr Asn Ser 225 230 235 240 Arg Phe Pro Val Pro Val Glu Met Met Tyr Thr Ala Arg Asn Glu Asn 245 250 255 Gln Val Val Gln Pro Gln Asn Gly Arg Val Thr Leu Asp Gly Glu Leu 260 265 270 Leu Gly Thr Thr Pro Leu Leu Ala Val Asn Ile Cys Lys Phe Lys Gly 275 280 285 Glu Val Ile Ala Lys Asn Gly Asp Val Arg Ser Tyr Arg Met Asp Met 290 295 300 Glu Ile Thr Asn Thr Asp Gly Thr Pro Ile Asp Pro Thr Glu Asp Thr 305 310 315 320 Pro Gly Pro Ile Gly Ser Pro Asp Phe Gln Gly Ile Leu Phe Gly Val 325 330 335 Ala Ser Gln Arg Asn Lys Asn Glu Gln Asn Pro Ala Thr Arg Ala His 340 345 350 Glu Ala Asn Ile Asn Thr Gly Gly Asp Gln Tyr Ala Pro Lys Leu Ala 355 360 365 Gln Val Lys Phe Phe Ser Glu Ser Gln Asp Phe Glu Val His Gln Pro 370 375 380 Thr Val Phe Thr Pro Val Gly Val Val Gly Asp Ser Ser His Pro Phe 385 390 395 400 Arg Gln Trp Val Leu Pro Arg Tyr Gly Gly His Leu Thr Asn Asn Thr 405 410 415 His Leu Ala Pro Ala Val Ala Pro Leu Phe Pro Gly Glu Gln Ile Leu 420 425 430 Phe Phe Arg Ser Gln Ile Pro Ser Ser Gly Gly His Glu Ser Gly Tyr 435 440 445 Val Asp Cys Leu Val Pro Gln Glu Trp Val Gln His Phe Tyr Gln Glu 450 455 460 Ala Ala Thr Ala Gln Ser Glu Val Ala Leu Ile Arg Phe Ile Asn Pro 465 470 475 480 Asp Thr Gly Arg Val Leu Phe Glu Ala Lys Leu His Lys Gln Gly Phe 485 490 495 Ile Thr Val Ala His Thr Gly Asp Asn Pro Ile Val Met Pro Pro Asn 500 505 510 Gly Tyr Phe Arg Phe Glu Ala Trp Val Asn Gln Phe Tyr Ser Leu Ala 515 520 525 Pro Val Gly Thr Gly Asn Gly Arg Arg Arg Ile Gln 530 535 540 <210> 7 <211> 535 <212> PRT <213> Norovirus <400> 7 Met Lys Met Ala Ser Asn Asp Ala Ala Pro Ser Asn Asp Gly Ala Ala 1 5 10 15 Gly Leu Val Pro Glu Val Asn Asn Glu Thr Met Ala Leu Glu Pro Val 20 25 30 Ala Gly Ala Ser Ile Ala Ala Pro Leu Thr Gly Gln Asn Asn Val Ile 35 40 45 Asp Pro Trp Ile Arg Leu Asn Phe Val Gln Ala Pro Asn Gly Glu Phe 50 55 60 Thr Val Ser Pro Arg Asn Ser Pro Gly Glu Val Leu Leu Asn Leu Glu 65 70 75 80 Leu Gly Pro Glu Leu Asn Pro Tyr Leu Ala His Leu Ser Arg Met Tyr 85 90 95 Asn Gly Tyr Ala Gly Gly Val Glu Val Gln Val Leu Leu Ala Gly Asn 100 105 110 Ala Phe Thr Ala Gly Lys Leu Val Phe Ala Ala Val Pro Pro His Phe 115 120 125 Pro Leu Glu Asn Ile Ser Pro Gly Gln Ile Thr Met Phe Pro His Val 130 135 140 Ile Ile Asp Val Arg Thr Leu Glu Pro Val Leu Leu Pro Leu Pro Asp 145 150 155 160 Val Arg Asn Asn Phe Phe His Tyr Asn Gln Gln Asn Glu Pro Arg Met 165 170 175 Arg Leu Val Ala Met Leu Tyr Thr Pro Leu Arg Ser Asn Gly Ser Gly 180 185 190 Asp Asp Val Phe Thr Val Ser Cys Arg Val Leu Thr Arg Pro Ser Pro 195 200 205 Asp Phe Asp Phe Asn Tyr Leu Val Pro Pro Thr Val Glu Ser Lys Thr 210 215 220 Lys Pro Phe Thr Leu Pro Ile Leu Thr Ile Gly Glu Leu Thr Asn Ser 225 230 235 240 Arg Phe Pro Val Pro Ile Asp Glu Leu Tyr Thr Ser Pro Asn Glu Ser 245 250 255 Leu Val Val Gln Pro Gln Asn Gly Arg Cys Ala Leu Asp Gly Glu Leu 260 265 270 Gln Gly Thr Thr Gln Leu Leu Pro Thr Ala Ile Cys Ser Phe Arg Gly 275 280 285 Arg Ile Asn Gln Lys Val Ser Gly Glu Asn His Val Trp Asn Met Gln 290 295 300 Ile Thr Asn Ile Asn Gly Thr Pro Phe Asp Pro Thr Glu Asp Val Pro 305 310 315 320 Ala Pro Leu Gly Thr Pro Asp Phe Ser Gly Lys Leu Phe Gly Val Leu 325 330 335 Ser Gln Arg Asp His Asp Asn Ala Cys Arg Ser His Asp Ala Val Ile 340 345 350 Ala Thr Asn Ser Ala Lys Phe Thr Pro Lys Leu Gly Ala Ile Gln Ile 355 360 365 Gly Thr Trp Glu Gln Asp Asp Val His Ile Asn Gln Pro Thr Lys Phe 370 375 380 Thr Pro Val Gly Leu Phe Glu Ser Glu Gly Phe Asn Gln Trp Thr Leu 385 390 395 400 Pro Asn Tyr Ser Gly Ala Leu Thr Leu Asn Met Gly Leu Ala Pro Pro 405 410 415 Val Ala Pro Thr Phe Pro Gly Glu Gln Ile Leu Phe Phe Arg Ser His 420 425 430 Ile Pro Leu Lys Gly Gly Val Ala Asp Pro Val Ile Asp Cys Leu Leu 435 440 445 Pro Gln Glu Trp Ile Gln His Leu Tyr Gln Glu Ser Ala Pro Ser Gln 450 455 460 Thr Asp Val Ala Leu Ile Arg Phe Thr Asn Pro Asp Thr Gly Arg Val 465 470 475 480 Leu Phe Glu Ala Lys Leu His Arg Ser Gly Tyr Ile Thr Val Ala Asn 485 490 495 Thr Gly Ser Arg Pro Ile Val Val Pro Ala Asn Gly Tyr Phe Arg Phe 500 505 510 Asp Ser Trp Val Asn Gln Phe Tyr Ser Leu Ala Pro Met Gly Thr Gly 515 520 525 Asn Gly Arg Arg Arg Val Gln 530 535 <210> 8 <211> 540 <212> PRT <213> Norovirus <400> 8 Met Lys Met Ala Ser Asn Asp Ala Ala Pro Ser Asn Asp Gly Ala Ala 1 5 10 15 Gly Leu Val Pro Glu Gly Asn Asn Glu Thr Leu Pro Leu Glu Pro Val 20 25 30 Ala Gly Ala Ala Ile Ala Ala Pro Val Thr Gly Gln Asn Asn Ile Ile 35 40 45 Asp Pro Trp Ile Arg Thr Asn Phe Val Gln Ala Pro Asn Gly Glu Phe 50 55 60 Thr Val Ser Pro Arg Asn Ser Pro Gly Glu Ile Leu Leu Asn Leu Glu 65 70 75 80 Leu Gly Pro Asp Leu Asn Pro Tyr Leu Ala His Leu Ser Arg Met Tyr 85 90 95 Asn Gly Tyr Ala Gly Gly Val Glu Val Gln Val Leu Leu Ala Gly Asn 100 105 110 Ala Phe Thr Ala Gly Lys Ile Leu Phe Ala Ala Val Pro Pro Asn Phe 115 120 125 Pro Val Glu Phe Leu Ser Pro Ala Gln Ile Thr Met Leu Pro His Leu 130 135 140 Ile Val Asp Val Arg Thr Leu Glu Pro Ile Met Ile Pro Leu Pro Asp 145 150 155 160 Val Arg Asn Thr Phe Phe His Tyr Ser Asn Gln Pro Asn Ser Arg Met 165 170 175 Arg Leu Val Ala Met Leu Tyr Thr Pro Leu Arg Ser Asn Gly Ser Gly 180 185 190 Asp Asp Val Phe Thr Val Ser Cys Arg Val Leu Thr Arg Pro Thr Pro 195 200 205 Asp Phe Glu Phe Thr Tyr Leu Val Pro Pro Ser Val Glu Ser Lys Thr 210 215 220 Lys Pro Phe Ser Leu Pro Ile Leu Thr Leu Ser Glu Leu Thr Asn Ser 225 230 235 240 Arg Phe Pro Val Pro Ile Asp Ser Leu Phe Thr Ala Gln Asn Asn Val 245 250 255 Leu Gln Val Gln Cys Gln Asn Gly Arg Cys Thr Leu Asp Gly Glu Leu 260 265 270 Gln Gly Thr Thr Gln Leu Leu Pro Ser Gly Ile Cys Ala Phe Arg Gly 275 280 285 Arg Val Thr Ala Gln Ile Asn Gln Arg Asp Arg Trp His Met Gln Leu 290 295 300 Gln Asn Leu Asn Gly Thr Thr Tyr Asp Pro Thr Asp Asp Val Pro Ala 305 310 315 320 Pro Leu Gly Thr Pro Asp Phe Lys Gly Val Val Phe Gly Met Val Ser 325 330 335 Gln Arg Asn Val Gly Asn Asp Ala Pro Gly Ser Thr Arg Ala Gln Gln 340 345 350 Ala Trp Val Ser Thr Tyr Ser Pro Gln Phe Val Pro Lys Leu Gly Ser 355 360 365 Val Asn Leu Arg Ile Ser Asp Asn Asp Asp Phe Gln Phe Gln Pro Thr 370 375 380 Lys Phe Thr Pro Val Gly Val Asn Asp Asp Asp Asp Gly His Pro Phe 385 390 395 400 Arg Gln Trp Glu Leu Pro Asn Tyr Ser Gly Glu Leu Thr Leu Asn Met 405 410 415 Asn Leu Ala Pro Pro Val Ala Pro Asn Phe Pro Gly Glu Gln Leu Leu 420 425 430 Phe Phe Arg Ser Phe Val Pro Cys Ser Gly Gly Tyr Asn Gln Gly Ile 435 440 445 Ile Asp Cys Leu Ile Pro Gln Glu Trp Ile Gln His Phe Tyr Gln Glu 450 455 460 Ser Ala Pro Ser Gln Ser Asp Val Ala Leu Ile Arg Tyr Val Asn Pro 465 470 475 480 Asp Thr Gly Arg Thr Leu Phe Glu Ala Lys Leu His Arg Ser Gly Tyr 485 490 495 Ile Thr Val Ala His Ser Gly Asp Tyr Pro Leu Val Val Pro Ala Asn 500 505 510 Gly His Phe Arg Phe Asp Ser Trp Val Asn Gln Phe Tyr Ser Leu Ala 515 520 525 Pro Met Gly Thr Gly Asn Gly Arg Arg Arg Ala Gln 530 535 540 <210> 9 <211> twenty one <212> DNA <213> Artificial Sequence <400> 9 tcgttcgttc gttcgttcgt t 21 <210> 10 <211> 25 <212> DNA <213> Artificial Sequence <400> 10 tcgttcgttc gttcgttcgt tcgtt 25 <210> 11 <211> twenty one <212> DNA <213> Artificial Sequence <400> 11 tcgtcgtcgt cgtcgtcgtc g 21

Claims

1. A norovirus virus-like particle or an active fragment thereof, said virus-like particle or active fragment thereof consisting of the amino acid sequence shown in SEQ ID NO:

5.

2. A composition for inducing an immune response to norovirus in animals, comprising norovirus virus-like particles or an active fragment thereof as described in claim 1.

3. The composition of claim 2, wherein the animal is a mammal.

4. The composition of claim 3, wherein the animal is a human.

5. The composition of claim 2, wherein the norovirus is GII.6 and / or GII.7 norovirus.

6. The composition of any one of claims 2 to 5, wherein the composition is a pharmaceutical composition for the prevention and / or treatment of norovirus infection.

7. The composition of any one of claims 2 to 5, wherein the composition is a vaccine for the prevention and / or treatment of norovirus infection.

8. The composition of claim 6, wherein the pharmaceutical composition further comprises one or more adjuvants selected from aluminum adjuvants, TRL adjuvants, and saponin adjuvants.

9. The composition of claim 8, wherein the aluminum adjuvant is selected from one or more of aluminum hydroxide, aluminum phosphate, and aluminum sulfate.

10. The composition of claim 9, wherein the aluminum adjuvant is aluminum hydroxide.

11. The composition of claim 8, wherein the TRL adjuvant is a TRL9 adjuvant.

12. The composition of claim 11, wherein the TRL9 adjuvant is a CPG adjuvant.

13. The composition of claim 12, wherein the nucleotide sequence of the CPG adjuvant is selected from one of SEQ ID NO: 9, SEQ ID NO: 10 or SEQ ID NO:

11.

14. The composition of claim 8, wherein the saponin adjuvant is QS21 or Iscom adjuvant.

15. The composition of claim 6, wherein the norovirus infection is gastroenteritis.

16. The composition of claim 6, wherein the norovirus infection is viral acute gastroenteritis.

17. An immunoassay kit comprising norovirus virus-like particles as described in claim 1 or an active fragment thereof, or a composition as described in any one of claims 2 to 16.

18. The immunoassay kit of claim 17, wherein the kit is a kit for detecting norovirus.

19. Use of the norovirus virus-like particles or active fragments thereof as claimed in claim 1, the composition as claimed in any one of claims 2 to 16, or the immunoassay kit as claimed in claim 17 or 18 in the preparation of the following products, (1) Medications used to prevent and / or treat norovirus infection; (2) A kit for diagnosing norovirus infection; or (4) Immunogens for developing norovirus antibodies.

20. The use according to claim 19, wherein the norovirus infection is gastroenteritis.

21. The use according to claim 19, wherein the norovirus infection is viral acute gastroenteritis.

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