KR20130128752A - Antibody of specific binding to na protein of influenza viruses - Google Patents

Abstract

The present invention relates to a binding molecule which binds to new influenza virus specifically. The new influenza virus-specific molecule is useful to prevent and treat diseases caused by the new influenza virus, and make a diagnosis of the new influenza virus. [Reference numerals] (AA) Influenza virus used in infecting cells

Description

Influenza virus NA protein specific antibody {Antibody of specific binding to NA protein of influenza viruses}

The present invention relates to antibodies that specifically bind to swine influenza virus NA proteins.

The causative virus of influenza is Orthomyxoviridae Family < / RTI > RNA (ssRNA) virus. Influenza viruses are usually divided into A, B, and C types. Influenza A virus infects not only humans but also birds and swine. Influenza C causes only mild illness in humans. Influenza A is subtyped again by surface antigens hemagglutanin (H) and neuramidase (N) antigens. H antigens are known to H1-H16, and N antigens are known to N1-N9. The gene for type A influenza is composed of 8 gene fragments, and for this reason, genetic recombination can occur.

Avian influenza viruses and swine influenza viruses do not infect humans well because of interspecies barriers. However, in rare cases infectious diseases exceed species barriers. In 1976, a case of infection with swine influenza (H1N1) occurred in New Jersey, USA. There were 13 infected patients and one of them died. In 1997 in Hong Kong, an infectious disease caused by avian influenza H5N1 occurred, and infection by the H5N1 influenza virus is still sporadic. By June 1, 2009, there were 432 cases of avian influenza in all over the world, of which 262 (60%) died.

Influenza A H1N1 has been identified as the causative microorganism of the new influenza A (H1N1) pdm09. However, unlike Influenza A (H1N1), which caused seasonal influenza, it became known that the causative virus of swine influenza [A (H1N1) pdm09] was caused by pigs and became known as S-OIV (swine influenza virus) ) The name changed to pdm09 virus. Molecular biology research has shown that the A (H1N1) pdm09 virus is a recombination of four influenza virus genes. The four influenza viruses are the North American swine influenza virus, the North American avian influenza virus, the human influenza virus and the Eurasian swine influenza virus, respectively.

Currently, the influenza virus diagnosis method is viral culture, and it is known that the sensitivity of the diagnosis is good when the culture is well matched (Reina, 1997). However, the virus culture method has a disadvantage in that it is costly in terms of experimental results, and it takes much time to deliver and confirm the virus to the laboratory.

As a commercialized rapid diagnosis reagent, there is a simultaneous detection kit of A-type circular and B-type rounds for human body, and a kit for detecting only A-type rounds in case of animals. However, the current commercialized influenza rapid diagnostic kit has a disadvantage that it can not distinguish the seasonal flu from the swine flu [A (H1N1) pdm09] virus which is currently popular. The current method for distinguishing between seasonal influenza and the new influenza A (H1N1) pdm09 virus is real-time polymerase chain reaction (RT-PCR) However, it has expensive equipment and high technical power, which makes it less convenient to use.

It is an object of the present invention to provide a binding molecule specifically binding to a swine influenza virus.

The present invention relates to a binding molecule that specifically binds to a NA protein of a new influenza virus, wherein the epitope to which the binding molecule binds relates to a binding molecule comprising P272 in the amino acid sequence of the NA protein. In one embodiment of the invention, the NA protein is SEQ ID NO: 1.

In one embodiment of the invention, the epitope to which the binding molecule binds is a binding molecule that specifically binds to the NA protein of the new influenza virus comprising P272 in the amino acid sequence of the NA protein; And a novel influenza virus therapeutic agent, and in one embodiment of the invention, the NA protein is SEQ ID NO: 1.

In one embodiment of the present invention, a vector comprising a gene encoding the binding molecule is provided.

In one embodiment of the present invention, a cell line comprising the vector is provided, and in one embodiment of the present invention, the cell line is F2N, HEK293, CHOK1, DG44, DXB11, CHO-S, BHK, Sp2 / 0, or NS.

In one embodiment of the present invention, there is provided a method of producing a binding molecule by transfecting the vector into a cell line.

In one embodiment of the present invention, providing a composition for preventing or treating a new influenza virus comprising the binding molecule, In one embodiment of the present invention, the composition for preventing or treating is 0.001 ~ 100 mg / kg To administer, in one embodiment of the present invention, the prophylactic or therapeutic composition is for rats, dogs, pigs, cattle, horses, rabbits, llamas, ferrets, and in one embodiment of the present invention, the prevention or Therapeutic compositions are in oral formulations, external preparations, pre-filled syringe solutions, suppositories, sterile injectable solutions or lyophilized forms.

In one embodiment of the present invention, there is provided a method for providing information on the presence of a new influenza virus comprising the step of reacting the binding molecule with the desired sample.

In one embodiment of the present invention, a diagnostic kit for a new influenza virus comprising said binding molecule.

The titer of the antibody acts to inhibit hemagglutination between influenza virus and erythrocytes by antibodies against HA1, the head portion of the HA protein of influenza virus produced by immunization and infection of influenza virus. It is measured using the hemagglutination inhibitory effect of the antibody.

The evasion mutation is that co-culture of the influenza virus used to make a monoclonal antibody with hemagglutination inhibitory ability with the monoclonal antibody results in a variant influenza virus that survives the inhibition of the monoclonal antibody, which is mainly a virus. It is called an evasion mutation because it has a mutation in the influenza virus surface antigen to which the monoclonal antibody acts and thus avoids the action of the monoclonal antibody. This avoidance mutation occurs in epitopes on which monoclonal antibodies act, and thus can be used to locate epitopes.

The transforming growth method used in the present invention can be carried out by a method known in the art, for example, but not limited to, by making a competent cell using CaCl ₂ buffer, and then introducing the expression vector into a host cell The transfection may be introduced into the host cell by transfection, transfection, transient transfection, microinjection, cell fusion, calcium phosphate precipitation, electroporation, or the like.

The novel influenza therapeutic agent used in the present invention may be, but is not limited to, Tamiflu or Relenza.

Since the antibody of the present invention specifically binds to the NA protein of the H1N1 influenza virus, it is useful as a method for treating the H1N1 virus and diagnosing infection.

1 is a result confirming the binding of the binding molecule mAb319 to the various viruses according to an embodiment of the present invention through immunofluorescence.

Hereinafter, the words of the present invention are defined as follows.

As used herein, "neuraminidase" (hereinafter referred to as "NA") is an envelope protein of influenza virus, and refers to an enzyme that hydrolyzes neuramic acid to separate sialic acid. NA is known to act when influenza virus flows out of a host cell.

Binding molecule " as used herein refers to an intact immunoglobulin comprising a monoclonal antibody, such as a chimeric, humanized or human monoclonal antibody, or an immunoglobulin that binds to an antigen, for example, Receptor or protein capable of binding to a variable domain or substrate comprising an immunoglobulin fragment competing with intact immunoglobulin for binding to influenza A virus monomer HA or trimer HA. Regardless of the structure, the antigen-binding fragment binds to the same antigen recognized by intact immunoglobulins. An antigen-binding fragment comprises two or more continuations of the amino acid sequence of a binding molecule, at least 20 contiguous amino acid residues, at least 25 contiguous amino acid residues, at least 30 contiguous amino acid residues, at least 35 contiguous amino acid residues, at least 40 contiguous amino acid residues. , At least 50 contiguous amino acid residues, at least 60 contiguous amino acid residues, at least 70 contiguous amino acid residues, at least 80 contiguous amino acid residues, at least 90 contiguous amino acid residues, at least 100 contiguous amino acid residues, at least 125 contiguous amino acid residues, Peptides or polypeptides comprising an amino acid sequence of at least 150 contiguous amino acid residues, at least 175 contiguous amino acid residues, at least 200 contiguous amino acid residues, or at least 250 contiguous amino acid residues. “Antigen-binding fragments” specifically include Fab, F (ab '), F (ab') ₂ , Fv, dAb, Fd, complementarity determining region (CDR) fragments, single-chain antibodies (scFv), bivalent Single-chain antibodies, single-chain phage antibodies, diabodies, triabodies, tetrabodies, polypeptides containing one or more fragments of immunoglobulin sufficient to bind a particular antigen to a polypeptide, and the like. The fragments may be produced synthetically or by enzymatic or chemical digestion of complete immunoglobulins or may be produced genetically by recombinant DNA techniques. Production methods are well known in the art.

As used herein, the term “pharmaceutically acceptable excipient” refers to an inert material that is combined into an active molecule, such as a drug, agent or binding molecule, to produce an acceptable or convenient dosage form. Pharmaceutically acceptable excipients are nontoxic or are excipients that are acceptable to the recipient for their intended use, at least in the doses and concentrations in which the toxicity is used, and with other components of the formulation including drugs, agents or binding powders. It is compatible.

As used herein, the term " therapeutically effective amount " refers to the amount of the binding molecule of the present invention effective for prevention or treatment before or after exposure of influenza A virus.

The composition containing the substance according to the present invention can be administered orally or parenterally in the form of powders, granules, tablets, capsules, oral liquid preparations such as suspensions, emulsions, syrups and aerosols, external preparations, pre-filled syringe solution, or lyophilized form. More specifically, when formulating the composition, it can be prepared using a diluent or an excipient such as a filler, an extender, a binder, a wetting agent, a disintegrant, a surfactant, and the like. Solid formulations for oral administration include tablets, pills, powders, granules, capsules and the like, which may contain at least one excipient such as starch, calcium carbonate, sucrose ), Lactose, gelatin and the like. In addition to simple excipients, lubricants such as magnesium stearate and talc may also be used. Examples of the liquid preparation for oral use include suspensions, solutions, emulsions, syrups and the like, and various excipients such as wetting agents, sweeteners, fragrances, preservatives, etc. in addition to commonly used diluents such as water and liquid paraffin . Formulations for parenteral administration include sterilized aqueous solutions, non-aqueous solutions, suspensions, emulsions, freeze-dried preparations and suppositories. Examples of the suspending agent include propylene glycol, polyethylene glycol, vegetable oil such as olive oil, injectable ester such as ethyl oleate, and the like. Examples of the suppository base include witepsol, macrogol, tween 61, cacao paper, laurin, glycerogelatin and the like.

The compounds of the present invention, particularly the binding molecules of the present invention, are particularly useful as diagnostic reagents in the detection of swine influenza virus.

Preferably, the compounds of the present invention used in the diagnostic composition are detectably labeled. Various methods that can be used to label biomolecules are well known to those skilled in the art and are contemplated within the scope of the present invention. These methods are described in Tijssen, 'Practice and theory of enzyme immunoassays', Burden, RH and von Knippenburg (Eds), Volume 15 (1985), 'Basic methods in molecular biology'; Davis LG, Dibmer MD; Acad. Press, London (1987), or in the series 'Methods in Enzymology', Academic Press, Inc .; Battey Elsevier (1990), Mayer et al., (Eds.) Immunochemical methods in cell and molecular biology.

There are many other markings and marking methods known to those skilled in the art. Examples of marking classes that can be used in the present invention include enzymes, radioactive isotopes, colloidal metals, fluorescent compounds, chemiluminescent compounds and bioluminescent compounds.

Commonly used markers include fluorescent substances (e.g., fluorescein, rhodamine, Texas red, etc.), enzymes (such as horseradish peroxidase,? -Galactosidase, alkaline phosphatase), radioactive isotopes 32 P or 125 I), biotin, digoxigenin, colloidal metals, chemiluminescent or bioluminescent compounds (such as dioxetane, luminol or acridinium). Methods of labeling enzymes or biotinyl groups such as covalent bonding, iodination, phosphorylation, biotinylation, and the like are well known in the art.

Detection methods include, but are not limited to, autoradiography, fluorescence microscopy, direct and indirect enzyme reactions, and the like. Commonly used detection assays include radioisotopes or non-radioisotope methods. These include Western blotting, overlay-analysis, Radioimmuno Assay (RIA) and Immune Radioimmunometric Assay (IRMA), Enzyme Immuno Assay (EIA), Enzyme Linked Immuno Sorbent Assay (ELISA), Fluorescent Immuno Assay (CIA) and Chemiluminoluminescent Immune Assay).

The antibody according to the present invention can be used in the form of a drug-conjugated antibody-drug conjugate. When an antibody-drug conjugate (ADC), that is, an immunoconjugate, is used for local delivery of a drug, the drug moiety can be targeted to the infected cells. If the drug is administered without conjugation, This can lead to unacceptable levels of toxicity. By increasing the selectivity of polyclonal and monoclonal antibodies (mAbs) as well as drug-linking and drug-releasing properties, the maximum efficacy and minimal toxicity of the ADC can be improved.

Conventional means of attaching the drug moiety to the antibody, ie, via covalent linkage, generally results in a heterogeneous molecular mixture in which the drug moiety is attached to numerous sites on the antibody. For example, cytotoxic drugs can typically be conjugated with an antibody, often through numerous lysine residues of the antibody, to produce a heterogeneous antibody-drug conjugate mixture. Depending on the reaction conditions, such heterogeneous mixtures typically have an antibody distribution between 0 and about 8 or more attached to the drug moiety. In addition, each aliquot of the conjugate with a specific integer ratio drug moiety-to-antibody is a potentially heterogeneous mixture in which the drug moiety is attached to various sites on the antibody. Antibodies are large, complex, structurally diverse biomolecules, and often have many reactive functional groups. Reactivity with linker reagents and drug-linker intermediates depends on factors such as pH, concentration, salt concentration and cosolvent.

Hereinafter, the present invention will be described in detail. However, the following examples are intended to illustrate the contents of the present invention and are not intended to limit the scope of the invention. The documents cited in the present invention are incorporated herein by reference.

Various embodiments described herein are described with reference to the drawings. In the following description, for purposes of complete understanding of the present invention, various specific details are set forth, such as specific forms, compositions, and processes, and the like. However, certain embodiments may be practiced without one or more of these specific details, or with other known methods and forms. In other instances, well-known processes and manufacturing techniques are not described in any detail, in order not to unnecessarily obscure the present invention.

Example

Example  1. Virus Concentration and Purification

Inactivated swine influenza virus culture (A / Korea / 01/2009) was provided. The method of S. Kodiballi et al. (1993) was used to concentrate and purify antigens from the newly isolated influenza virus (A / Korea / 01/2009, A (H1N1) pdm09).

Specifically, influenza virus incubated with about 2 ⁵ NA activity was inactivated with β-propiolactone and subjected to primary centrifugation at 4,000 xg for 30 minutes, and then the supernatant was collected at 70,000 xg at 3 Ultracentrifugation (L8-70M, Beckman, USA) for hours. The centrifuged pellet was resuspended in sucrose (1/25 times the amount of sucrose), sucrose density gradient ultracentrifugation (100,000 xg, 90 min) And then the fraction showing hemagglutination activity was isolated. To this concentrated virus fraction, 5% (w / v) n-octyl-bD-gluco pyranoside was added and the virus was disrupted by treatment at 22 ° C for 2 hours . Thereafter, the pellet was centrifuged at 70,000 xg for 90 minutes, the pellet was dissolved in 0.1 M NaCl, and centrifuged again at 10,000 xg for 30 minutes to isolate the influenza virus.

Example  2. Monoclonal antibody production

2.1 Immunogen preparation

The immunogen was mixed with complete Freund's adjuvant (Sigma) and emulsified sufficiently. The mice were inoculated 3 times at 1 week intervals with 200 ㎍ each, and the tail vein injection was performed 3 times.

2.2 Serum collection, Potency  Measurement and cell fusion

A small amount of blood was collected from the tail of the immunized mouse (Balb / c), and the serum was separated, and the activity was measured by enzyme immunoassay. Antibody titers were adsorbed on an ELISA plate, and antiserum was diluted 10, 100, 1,000, and 10,000 times with 10 times dilution. Secondary reactions were performed using goat anti-mouse IgG-peroxidase (Goat anti-mouse IgG-peroxidase) and developed with TMB. Cell fusion was performed when the absorbance of the enzyme was about 3 times higher than the absorbance of normal mouse serum as a cutoff and the titer of the antiserum was more than 1,000 times and the titer was more than the cutoff (Table 1).

A novel influenza-specific hemagglutinin antibody in a hybrid cell line that isolates splenocytes of immunized mice and conjugates them with myeloma cells (SP2 / 0) to secrete monoclonal antibodies through Hemagglutinin Inhibition Assay Were selected and several single cell populations were established by the limiting dilution method. Cell lines were cultured in large quantities to inoculate cells in the abdominal cavity of Balb / c mice to obtain ascites containing a large amount of antibody, and to purify IgG antibodies using protein G gel.

Dilution of antiserum dilution 10 times 100 times 1,000 times 10,000 times Normal mouse serum
Absorbance ( OD ₄₅₀ ) 0.091 0.095 0.110 0.096 Immune mouse serum
Absorbance ( OD ₄₅₀ ) 1.820 1.026 0.450 0.201

2.3 Mouse monoclonal swine influenza virus Hemiglutinin  Antibody Screening

30 μl of PBS (phosphate buffered saline) was added to each well of the U-plated. 30 μl of the sample was mixed with the first well and diluted by 1/2 with the next well. After diluting 30 μl with the final well, And the control group did not contain any specimen. The virus was diluted to 2 ² HAU (hemagglutination unit) in various types of suntypes and 30 μl of each sample was added to the well. In the control, 2 ² HA virus was added to the first well and diluted 3 times with 1/2 Respectively.

After standing at room temperature for 30 minutes, 30 μl of 1% chicken erythrocytes were dispensed into each well, and erythrocytes were similarly dispensed to the control group. Again, the cells were allowed to stand at room temperature for 40 minutes, and cell lines in which erythrocyte sedimentation occurred were selected as compared with the control group.

2.4 Mouse Monoclonal  Antibody purification

A large number of cell strains selected in Example 2.3 were cultured and inoculated into peritoneal cavity of Balb / c mice to obtain a plurality of antibodies containing a large amount of antibody, and then the plurality was centrifuged at 2500 rpm for 10 minutes and the precipitate was removed. The plurality was diluted with the above-mentioned 4-fold volumes of binding buffer and allowed to stand at room temperature for 1 hour.

The diluted solution was centrifuged at 12000 rpm at 4 캜 for 5 minutes (High speed centrifugation) and the precipitate was removed and the protein G gel was washed with 1.5M to 3M NaSCN in OD ₂₈₀ Lt; RTI ID = 0.0 > 0. < / RTI > The washed protein G gel was allowed to equilibrate with binding buffer. The prepared specimen was loaded and washed with binding buffer until the OD ₂₈₀ value of the washing solution was less than 0.05. Elution was carried out by elution buffer (2 times repeated), and after standing for 10 minutes, OD ₂₈₀ value of each fraction was measured.

A The measured OD ₂₈₀ value of 0.9 or more tubes only PD-10 in PBS (pH7.2) 0.1% NaN ₃ after dialysis, filtering the dialyzed antibody as a filter having a pore size of 0.2 ㎛, by measuring the OD ₂₈₀ value of the concentration .

Example  3. Swine influenza virus [A ( H1N1 ) pdm09 ] Binding characteristics of specific antibodies

In order to confirm whether the monoclonal antibody (mAbI319) obtained in Example 2 specifically binds to A (H1N1) pdm09, hemagglutinin inhibition (HAI) assay was performed for hemagglutinin inhibitory activity. The experiment was performed in the same manner as conventionally known, wherein ^a -pdmH1N1 H1N1 and ^b is Ferret antisera obtained from NIID (National Institute of Infectious Diseases in Japnan) for each of the A / Korea / 01/2009 and A (H1N1) in pdm09 ( ferret antiserum) control sheep antiserum. On the other hand, anti-IFN-B ^c and anti-IFN-B ^d are the influenza B reference antiserum against B / brisbane / 60/2008 and B / Florida / 4/2006, respectively.

As a result, as shown in Table 2 below, mAbI319 antibody against other influenza subtypes showed no binding activity to H1N1, H3N2, B (Brisbane) and B (Florida) except for H1N1pdm, And it was found.

virus Antibody or anti-serum mAb I319 anti-pDM H1N1 ^a anti-PDM H1N1 ^b anti-H1N1 anti-H3N2 anti-IFN-B ^c anti-IFN-B ^d H1N1pdm 128 1280 2560 - - - - H1N1 - - - 640 - - - H3N2 - - - - 2560 - - B (Brisbane) - - - - 40 2560 80 B
(Florida) - - - - 80 40 2560

Example  4. Antigen Epitope  Specific: evasion mutation

After infecting MDCK cells with influenza virus, staining with an anti-NP antibody specific for mAbI319 influenza virus nucleoprotein, as shown in FIG. ), mI319 (19) and mI319 (22) influenza viruses did not bind mAbI319. Blue is MDCK nuclei stained by DAPI, green is stained by mAbI319 and anti-NP.

From this, it was found that escape mutant occurred at the site to which the monoclonal antibody mAbI319 binds, and the surface antigens (NA, HA, and M1319 (18), mI319 (19) and mI319 (22)) are generated. The base sequence of M2) was analyzed to find the site of avoidance mutation compared to wild type (KR01 or KR02). It can be seen that the site where the avoidance mutation occurred is the mAbI319 epitope. Sequence check results are shown in Table 3 below.

Wild virus and
escape mutants ^a HA ^b NA ^b IFA ^c 147 212 240 272 * 319 mAbI319 anti-NP KR02 (2-5) K E R P S O O mI319 (18) K E R L S X O mI319 (19) Q E R L S X O mI319 (22) * K E R L S X O KR01 K A Q P R O O

Analysis of the nucleotide sequence of the evasion mutations and comparison with that of the wild-type virus revealed that the amino acid NA 272 is an epitope of mABI319.

<110> Republic of Korea <120> Antibody of specific binding to NA protein of influenza viruses <130> IPDB46124 <160> 1 <170> Kopatentin 2.0 <210> 1 <211> 469 <212> PRT <213> A / Korea / 01/2009 segment 6 <400> 1 Met Asn Pro Asn Gln Lys Ile Ile Thr Ile Gly Ser Val Cys Met Thr   1 5 10 15 Ile Gly Met Ale Asn Ile Ile Leu Gln Ile Gly Asn Ile Ile Ser Ile              20 25 30 Trp Ile Ser His Ser Ile Gln Leu Gly Asn Gln Asn Gln Ile Glu Thr          35 40 45 Cys Asn Gln Ser Val Ile Thr Tyr Glu Asn Asn Thr Trp Val Asn Gln      50 55 60 Thr Tyr Val Asn Ile Ser Asn Thr Asn Phe Ala Gly Gln Ser Val  65 70 75 80 Val Ser Val Lys Leu Ala Gly Asn Ser Ser Leu Cys Pro Val Ser Gly                  85 90 95 Trp Ala Ile Tyr Ser Lys Asp Asn Ser Ile Arg Ile Gly Ser Lys Gly             100 105 110 Asp Val Phe Val Ile Arg Glu Pro Phe Ile Ser Cys Ser Pro Leu Glu         115 120 125 Cys Arg Thr Phe Phe Leu Thr Gln Gly Ala Leu Leu Asn Asp Lys His     130 135 140 Ser Asn Gly Thr Ile Lys Asp Arg Ser Ser Tyr Arg Thr Leu Met Ser 145 150 155 160 Cys Pro Ile Gly Glu Val Pro Ser Pro Tyr Asn Ser Arg Phe Glu Ser                 165 170 175 Val Ala Trp Ser Ala Ser Ala Cys His Asp Gly Ile Asn Trp Leu Thr             180 185 190 Ile Gly Ile Ser Gly Pro Asp Asn Gly Ala Val Ala Val Leu Lys Tyr         195 200 205 Asn Gly Ile Ile Thr Asp Thr Ile Lys Ser Trp Arg Asn Asn Ile Leu     210 215 220 Arg Thr Gln Glu Ser Glu Cys Ala Cys Val Asn Gly Ser Cys Phe Thr 225 230 235 240 Val Met Thr Asp Gly Pro Ser Asn Gly Gln Ala Ser Tyr Lys Ile Phe                 245 250 255 Arg Ile Glu Lys Gly Lys Ile Val Lys Ser Val Glu Met Asn Ala Pro             260 265 270 Asn Tyr His Tyr Glu Glu Cys Ser Cys Tyr Pro Asp Ser Ser Glu Ile         275 280 285 Thr Cys Val Cys Arg Asp Asn Trp His Gly Ser Asn Arg Pro Trp Val     290 295 300 Ser Phe Asn Gln Asn Leu Glu Tyr Gln Ile Gly Tyr Ile Cys Ser Gly 305 310 315 320 Ile Phe Gly Asp Asn Pro Arg Pro Asn Asp Lys Thr Gly Ser Cys Gly                 325 330 335 Pro Val Ser Ser Asn Gly Ala Asn Gly Val Lys Gly Phe Ser Phe Lys             340 345 350 Tyr Gly Asn Gly Val Trp Ile Gly Arg Thr Lys Ser Ile Ser Ser Arg         355 360 365 Asn Gly Phe Glu Met Ile Trp Asp Pro Asn Gly Trp Thr Gly Thr Asp     370 375 380 Asn Asn Phe Ser Ile Lys Gln Asp Ile Val Gly Ile Asn Glu Trp Ser 385 390 395 400 Gly Tyr Ser Gly Ser Phe Val Gln His Pro Glu Leu Thr Gly Leu Asp                 405 410 415 Cys Ile Arg Pro Cys Phe Trp Val Glu Leu Ile Arg Gly Arg Pro Lys             420 425 430 Glu Asn Thr Ile Trp Thr Ser Gly Ser Ser Ile Ser Phe Cys Gly Val         435 440 445 Asn Ser Asp Thr Val Gly Trp Ser Trp Pro Asp Gly Ala Glu Leu Pro     450 455 460 Phe Thr Ile Asp Lys 465

Claims (15)

In a binding molecule that specifically binds to NA protein of swine influenza virus,
The epitope to which the binding molecule binds is a binding molecule comprising P272 of the amino acid sequence of NA protein.
The method of claim 1,
The NA protein is a binding molecule, characterized in that SEQ ID NO: 1.
The epitope to which the binding molecule binds is a binding molecule that specifically binds to the NA protein of the new influenza virus including P272 in the amino acid sequence of the NA protein; And antibody-drug conjugates comprising the new influenza virus therapeutic.
The method of claim 3, wherein
The NA protein is a binding molecule, characterized in that SEQ ID NO: 1.
A vector comprising a gene encoding the binding molecule of claim 1.
A cell line comprising the vector of claim 5.
The method according to claim 6,
Wherein the cell line is F2N, HEK293, CHOK1, DG44, DXB11, CHO-S, BHK, Sp2 / 0 or NS.
A method of producing a binding molecule by transfecting a cell of claim 5 into a cell line.
The method of claim 8, wherein the cell line is F2N, HEK293, CHOK1, DG44, DXB11, CHO-S, BHK, Sp2 / 0, or NS.
A novel influenza virus prevention or treatment composition comprising the binding molecule of claim 1 or 2.
The method of claim 10,
The prophylactic or therapeutic composition is a swine influenza virus prevention or treatment composition, characterized in that administering 0.001 ~ 100 mg / kg.
The method of claim 10,
The preventive or therapeutic composition is a new influenza virus prevention or treatment composition, characterized in that for rats, dogs, pigs, cattle, horses, rabbits, llama or ferret.
The method of claim 10,
The prophylactic or therapeutic composition is an oral dosage form, an external preparation, a pre-filled syringe solution, a suppository, a sterile injectable solution or a lyophilized form for the prevention or treatment of the new influenza virus. Composition.
A method of providing information on the presence of a new influenza virus comprising the step of reacting the binding molecule of claim 1 with a target sample.
A diagnostic kit for swine flu virus comprising the binding molecule of claim 1.

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