WO2023167317A1 - ANTICORPS ANTI-SARS-CoV-2 DIRIGÉ CONTRE L'ANTIGÈNE DU SARS-CoV-2 DANS UN LIQUIDE CORPOREL, COMPRENANT UN MUTANT; PROCÉDÉ DE DÉTECTION DU SARS-CoV-2 À L'AIDE D'UN ANTICORPS; ET KIT CONTENANT UN ANTICORPS - Google Patents
ANTICORPS ANTI-SARS-CoV-2 DIRIGÉ CONTRE L'ANTIGÈNE DU SARS-CoV-2 DANS UN LIQUIDE CORPOREL, COMPRENANT UN MUTANT; PROCÉDÉ DE DÉTECTION DU SARS-CoV-2 À L'AIDE D'UN ANTICORPS; ET KIT CONTENANT UN ANTICORPS Download PDFInfo
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Classifications
-
- C—CHEMISTRY; METALLURGY
- C07—ORGANIC CHEMISTRY
- C07K—PEPTIDES
- C07K16/00—Immunoglobulins [IGs], e.g. monoclonal or polyclonal antibodies
- C07K16/08—Immunoglobulins [IGs], e.g. monoclonal or polyclonal antibodies against material from viruses
- C07K16/10—Immunoglobulins [IGs], e.g. monoclonal or polyclonal antibodies against material from viruses from RNA viruses
-
- G—PHYSICS
- G01—MEASURING; TESTING
- G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
- G01N33/00—Investigating or analysing materials by specific methods not covered by groups G01N1/00 - G01N31/00
- G01N33/48—Biological material, e.g. blood, urine; Haemocytometers
- G01N33/50—Chemical analysis of biological material, e.g. blood, urine; Testing involving biospecific ligand binding methods; Immunological testing
- G01N33/53—Immunoassay; Biospecific binding assay; Materials therefor
- G01N33/531—Production of immunochemical test materials
-
- G—PHYSICS
- G01—MEASURING; TESTING
- G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
- G01N33/00—Investigating or analysing materials by specific methods not covered by groups G01N1/00 - G01N31/00
- G01N33/48—Biological material, e.g. blood, urine; Haemocytometers
- G01N33/50—Chemical analysis of biological material, e.g. blood, urine; Testing involving biospecific ligand binding methods; Immunological testing
- G01N33/53—Immunoassay; Biospecific binding assay; Materials therefor
- G01N33/543—Immunoassay; Biospecific binding assay; Materials therefor with an insoluble carrier for immobilising immunochemicals
-
- G—PHYSICS
- G01—MEASURING; TESTING
- G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
- G01N33/00—Investigating or analysing materials by specific methods not covered by groups G01N1/00 - G01N31/00
- G01N33/48—Biological material, e.g. blood, urine; Haemocytometers
- G01N33/50—Chemical analysis of biological material, e.g. blood, urine; Testing involving biospecific ligand binding methods; Immunological testing
- G01N33/53—Immunoassay; Biospecific binding assay; Materials therefor
- G01N33/569—Immunoassay; Biospecific binding assay; Materials therefor for microorganisms, e.g. protozoa, bacteria, viruses
-
- C—CHEMISTRY; METALLURGY
- C07—ORGANIC CHEMISTRY
- C07K—PEPTIDES
- C07K14/00—Peptides having more than 20 amino acids; Gastrins; Somatostatins; Melanotropins; Derivatives thereof
- C07K14/005—Peptides having more than 20 amino acids; Gastrins; Somatostatins; Melanotropins; Derivatives thereof from viruses
- C07K14/08—RNA viruses
- C07K14/165—Coronaviridae, e.g. avian infectious bronchitis virus
Definitions
- the present invention relates to anti-SARS-CoV-2 antibodies against SARS-CoV-2 antigens in body fluids, including mutant strains, methods for detecting SARS-CoV-2 using the antibodies, kits containing the antibodies, and the like. .
- a method for detecting a protein that constitutes a SARS-CoV-2-derived nucleocapsid from a sample wherein the sample is the antibody or fragment thereof according to any one of [1] to [6], or a combination thereof The above method, comprising the step of contacting.
- the method of [7] wherein the sample is a biological sample.
- the kit of [11] which is in the form of an immunochromatographic strip.
- FIG. 8 shows the results of detection by sandwich ELISA using cultured virus-derived NP of each mutant strain.
- FIG. 9 shows SDS-PAGE and CBB staining of NPs mixed with saliva specimens.
- Left is a sample of raw saliva mixed with NP or PBS
- Light is a sample of heat-treated saliva mixed with NP or PBS.
- FIG. 10 shows detection by Western blot using each antibody of NP mixed with a saliva sample.
- FIG. 11 shows the results of detection by sandwich ELISA using each antibody for NP added to a saliva sample.
- the epitope recognized by the antibody of the present invention, a fragment thereof, or a combination thereof is the amino acid sequence represented by SEQ ID NO: 1 to 50, 51 to 105, 106 to 160, or 161 to It is in the 207th position.
- the antibodies of the invention, or fragments thereof, or combinations thereof specifically bind to proteins that make up the SARS-CoV-2-derived nucleocapsid.
- An antibody of the invention is, for example, a complete immunoglobulin molecule, preferably IgM, IgD, IgE, IgA or IgG, more preferably IgG1, IgG2a, IgG2b, IgG3 or IgG4.
- Antibodies of the present invention can also be modified and/or altered antibodies, such as chimeric and humanized antibodies.
- Antibodies of the invention can also be monoclonal or polyclonal antibodies, modified or altered monoclonal or polyclonal antibodies, or recombinantly or synthetically produced or synthesized antibodies.
- the antibody of the present invention is preferably a monoclonal antibody from the viewpoint of accurate, convenient, and rapid detection of SARS-CoV-2-derived NPs.
- Antibodies or fragments thereof, or combinations thereof of the present invention may also be antibody derivatives such as bifunctional antibodies, or antibody constructs such as single-chain Fvs (scFv), bispecific scFvs or antibody fusion proteins. obtain.
- scFv single-chain Fvs
- scFv single-chain Fvs
- a combination of fragments although the position of the epitope is not particularly limited, 1 to 207, especially 1 to 50, 51 to 105, 106 to 160 on the N-terminal side of the amino acid sequence represented by SEQ ID NO: 1 , or a combination of two or more antibodies or fragments that recognize epitopes 161 to 207, or 1 to 207 on the N-terminal side of the amino acid sequence represented by SEQ ID NO: 1, especially 1 to 50, 51 to 105
- a combination of an antibody or fragment that recognizes the 106th, 106th to 160th, or 161st to 207th epitopes and an existing antibody or fragment that recognizes other epitopes is preferred.
- the antibody of the present invention may specifically bind to a protein that constitutes a SARS-CoV-2-derived nucleocapsid.
- Specifically binding means in a general sense having a high binding affinity compared to other proteins, and in a preferred form, for example compared to the NPs of SARS, MERS, the binding affinity may mean that the
- the method of the present invention detects a protein that constitutes a SARS-CoV-2-derived nucleocapsid by at least one immunological test selected from the group consisting of ELISA, CLEIA, immunochromatography, and filter antigen assay. further comprising the step of
- the method of the present invention comprises the step of conducting an antigen-antibody reaction between an antibody of the present invention or a fragment thereof, or a combination thereof and SARS-CoV-2-derived NPs. In one aspect, the method of the invention comprises multiple steps of conducting an antigen-antibody reaction between an antibody of the invention, or a fragment thereof, or a combination thereof, and SARS-CoV-2-derived NPs. In one embodiment, the antibodies of the invention or fragments thereof, or combinations thereof, differ in epitope each time through the steps. In one aspect, an antibody or fragment thereof, or combination thereof, of the present invention is labeled with a labeling substance.
- the method of the present invention includes a step of performing an antigen-antibody reaction between an unlabeled antibody of the present invention or a fragment thereof, or a combination thereof and SARS-CoV-2-derived NPs, and labeled with a labeling substance.
- Antigen-antibody reaction between the antibody of the present invention or a fragment thereof, or a combination thereof (the epitope is different from the unlabeled antibody of the present invention, a fragment thereof, or a combination thereof) and SARS-CoV-2-derived NPs including the step of performing
- the method of the present invention includes the step of conducting an antigen-antibody reaction between the antibody of the present invention or a fragment thereof, or a combination thereof and SARS-CoV-2-derived NPs, and/or the antibody of the present invention or It further comprises the step of conducting an antigen-antibody reaction between a fragment thereof, or a combination thereof, and an antibody or a fragment thereof, or a combination thereof, which recognizes the antibody, a fragment thereof, or a combination thereof.
- the antibody of the present invention or a fragment thereof, or a combination thereof, and/or an antibody or a fragment thereof, or a combination thereof that recognizes the antibody or a fragment thereof, or a combination thereof is labeled with a labeling substance.
- sample is any sample that can contain SARS-CoV-2-derived NPs, and is not particularly limited, but includes, for example, biological samples.
- sample is used interchangeably with “specimen.”
- biological sample refers to human or animal blood, plasma, serum, urine, semen, cerebrospinal fluid, saliva, oral mucosa, nasal discharge, sweat, tears, ascites, amniotic fluid, feces, organs such as blood vessels or liver Any sample that can contain SARS-CoV-2-derived NPs, such as , tissues, cells, or extracts thereof, preferably from oral cavity, tonsil, nasal cavity, pharynx, larynx, trachea that is easy to collect.
- Non-Patent Document 6 put 1 to 2 ml of serum in a plastic tube that can be sealed after separation, cap it, and seal it with Parafilm. It may contain a coagulant, and if a blood collection tube containing a serum separator is used, transfer 1 to 2 ml of the serum after centrifugation into a plastic tube (preferably a sterilized tube), cap it, and seal it with Parafilm. .
- Non-Patent Document 6 whole blood was collected in a blood collection tube containing an anticoagulant (EDTA-Na or K), and 1 to 2 ml was dispensed into a plastic tube that can be sealed and capped. Then seal with Parafilm. If possible, blood cells are separated and peripheral blood mononuclear cells are suspended in a cell preservation medium and cryopreserved. Isolation of peripheral blood mononuclear cells is facilitated using BD Vacutainer® CPTTM mononuclear cell isolation blood collection tubes. In addition, when it is not possible to perform dispensing or blood cell separation after blood collection, blood may be collected using a PAXgene (registered trademark) RNA collection tube and stored frozen as it is.
- PAXgene registered trademark
- Non-Patent Document 6 sputum is collected when sputum is produced. If on mechanical ventilation, tracheal aspirate should be obtained using a sterile catheter under aseptic technique. Consider collecting bronchoalveolar lavage if not clinically contraindicated. The collected sputum or aspirate is placed in a plastic tube with a screw cap, capped and sealed with Parafilm.
- a nasopharyngeal swab is prepared by inserting a sterile cotton swab (thin swab for nasal cavity, such as a floss swab or a cotton swab containing rayon or polyester in the material) from the nostril, and thoroughly cleaning the nasopharynx. Place wipes and swabs into sterile spitz tubes containing 1-3 ml of viral transport fluid (VTM/UTM), cap and seal with Parafilm. If no virus transport solution is available, use PBS, physiological saline, or the like.
- VTM/UTM viral transport fluid
- Non-Patent Document 6 put 1 to 2 ml of saliva in a sterilized plastic tube, cap it, and seal it with Parafilm.
- a saliva collection kit such as Salivet (registered trademark) is used to suck saliva into cotton, and then the saliva is separated and collected by centrifugation.
- Non-Patent Document 6 1 to 2 ml of urine is placed in a test tube (Falcon tube, etc.), capped, and sealed with Parafilm.
- a test tube Fecon tube, etc.
- Parafilm For example, about 0.1 g (adzuki bean size) of stool is collected in a hermetically sealed plastic tube, capped, and then sealed with Parafilm.
- the “labeling substance” refers to the antibody of the present invention or a fragment thereof, or a combination thereof, and/or an antibody or a fragment thereof, or a combination thereof that recognizes the antibody or a fragment thereof, or a combination thereof. Any substance capable of detecting an antigen-antibody reaction is not particularly limited. From the viewpoint of accurate, convenient, and rapid detection of SARS-CoV-2-derived NPs, the labeling substance is one or more selected from enzymes, dyes, colloidal metal particles, latex particles, cellulose particles, and the like. preferable.
- Antibodies that recognize the antibodies of the invention or fragments thereof, or combinations thereof can also be modified and/or altered antibodies, such as chimeric and humanized antibodies.
- Antibodies that recognize the antibodies of the present invention or fragments thereof, or combinations thereof may also be monoclonal or polyclonal antibodies, modified or altered monoclonal or polyclonal antibodies, or recombinantly or synthetically produced or synthesized antibodies. .
- An antibody of the invention or a fragment thereof, or a fragment of an antibody that recognizes a combination thereof can be a Fab fragment or part of such immunoglobulin molecules, such as the VL-, VH- or CDR-regions, although the antibody of the invention Or a fragment thereof, or a fragment of an antibody that recognizes a combination thereof, is also as specific for an antibody of the invention, or a fragment thereof, or a combination thereof, as an antibody that recognizes an antibody of the invention, or a fragment thereof, or a combination thereof. can be recognized and bound to each other.
- Fragments of antibodies that recognize antibodies or fragments thereof, or combinations thereof, of the present invention include antibody fragments and separate light and heavy chains, Fab, Fab/c, Fv, Fab', F(ab') 2 and so on.
- a fragment of an antibody that recognizes an antibody of the invention or a fragment thereof, or a combination thereof is a Fab, Fab/c, Fv, Fab' or F(ab') 2 fragment.
- the antibody or fragment thereof of the present invention may be an antibody derivative such as a bifunctional antibody, or a single-chain Fv (scFv), bispecific It may be an antibody construct such as a sex scFv or an antibody fusion protein.
- an antibody derivative such as a bifunctional antibody, or a single-chain Fv (scFv), bispecific It may be an antibody construct such as a sex scFv or an antibody fusion protein.
- SARS-CoV-2-derived NPs are detected by any immunological test that can detect an antigen-antibody reaction between the antibody of the present invention or a fragment thereof, or a combination thereof and SARS-CoV-2-derived NPs. .
- Another aspect of the present invention is a method for diagnosing SARS-CoV-2 infection comprising the step of contacting a sample with an antibody or fragment thereof of the present invention, or a combination thereof (hereinafter referred to as "diagnostic method of the present invention"). (as may be the case).
- the diagnostic method of the present invention identifies proteins that make up the SARS-CoV-2-derived nucleocapsid by at least one immunological test selected from the group consisting of ELISA, CLEIA, immunochromatography, and filter antigen assay. Further comprising the step of detecting.
- the diagnostic method of the present invention includes a step of performing an antigen-antibody reaction between the unlabeled antibody of the present invention or a fragment thereof, or a combination thereof and SARS-CoV-2-derived NPs, and labeled with a labeling substance.
- Antigen antibody between the antibody of the present invention or a fragment thereof, or a combination thereof (the epitope is different from the unlabeled antibody of the present invention or a fragment thereof, or a combination thereof) and the SARS-CoV-2-derived NP including the step of conducting the reaction.
- the diagnostic method of the present invention includes the step of conducting an antigen-antibody reaction between the antibody of the present invention or a fragment thereof, or a combination thereof and SARS-CoV-2-derived NPs, and/or the antibody of the present invention. or a fragment thereof, or a combination thereof, and an antibody or fragment thereof, or a combination thereof, which recognizes the antibody, a fragment thereof, or a combination thereof, and a step of conducting an antigen-antibody reaction.
- the antibody of the present invention or a fragment thereof, or a combination thereof, and/or an antibody or a fragment thereof, or a combination thereof that recognizes the antibody or a fragment thereof, or a combination thereof is labeled with a labeling substance.
- the DNA sequence represented by SEQ ID NO: 4 was artificially synthesized, and by PCR using the DNA sequence as a template, the amino acid sequence represented by SEQ ID NO: 1 corresponded to the 1st to 207th regions (Fig. 1). A region from 1 to 621 of the DNA sequence represented by SEQ ID NO:4 was amplified.
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Abstract
La présente invention aborde le problème de la fourniture d'un moyen hautement sensible et hautement spécifique pour détecter une protéine constituant une nucléocapside dérivée du SARS-CoV-2. La présente invention concerne à titre d'exemple, un anticorps se liant à une protéine constituant la nucléocapside dérivée du SARS-CoV-2 ou un fragment de celui-ci, ou une combinaison de ceux-ci; un procédé de détection, à partir d'un échantillon, d'une protéine constituant la nucléocapside dérivée du SARS-CoV-2 à l'aide dudit anticorps ou d'un fragment de celui-ci, ou d'une combinaison de ceux-ci; et un kit de détection d'une protéine constituant la nucléocapside dérivée du SARS-CoV-2, le kit contenant ledit anticorps ou un fragment de celui-ci, ou une combinaison de ceux-ci.
Applications Claiming Priority (2)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| JP2022-033238 | 2022-03-04 | ||
| JP2022033238A JP2023128698A (ja) | 2022-03-04 | 2022-03-04 | 変異株を含む、体液中のＳＡＲＳ－ＣｏＶ－２抗原に対する抗ＳＡＲＳ－ＣｏＶ－２抗体、該抗体を用いてＳＡＲＳ－ＣｏＶ－２を検出する方法、および該抗体を含むキット |
Publications (1)
| Publication Number | Publication Date |
|---|---|
| WO2023167317A1 true WO2023167317A1 (fr) | 2023-09-07 |
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| Application Number | Title | Priority Date | Filing Date |
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| PCT/JP2023/008064 WO2023167317A1 (fr) | 2022-03-04 | 2023-03-03 | ANTICORPS ANTI-SARS-CoV-2 DIRIGÉ CONTRE L'ANTIGÈNE DU SARS-CoV-2 DANS UN LIQUIDE CORPOREL, COMPRENANT UN MUTANT; PROCÉDÉ DE DÉTECTION DU SARS-CoV-2 À L'AIDE D'UN ANTICORPS; ET KIT CONTENANT UN ANTICORPS |
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| JP (1) | JP2023128698A (fr) |
| WO (1) | WO2023167317A1 (fr) |
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| CN111647053A (zh) * | 2020-04-16 | 2020-09-11 | 军事科学院军事医学研究院生命组学研究所 | 多肽及其在新型冠状病毒检测、抗体或疫苗筛选中的应用 |
| CN113234145A (zh) * | 2020-08-12 | 2021-08-10 | 中山大学附属第五医院 | 特异性结合新型冠状病毒的抗体 |
| WO2021181994A1 (fr) * | 2020-03-10 | 2021-09-16 | デンカ株式会社 | Épitope d'anticorps dirigé contre la protéine structurelle du sras-cov-2, anticorps réagissant avec l'épitope, procédé de détection de sras-cov-2 à l'aide d'anticorps, kit de détection pour anticorps contenant sras-cov-2, procédé de détection d'anticorps anti-sras-cov-2 contenant un polypeptide d'épitope, kit de détection pour un anticorps anti-sras-cov-2 contenant un polypeptide d'épitope, vaccin contre sras-cov-2 contenant un polypeptide d'épitope, et agent thérapeutique pour une infection à sras-cov-2 contenant l'anticorps |
| WO2022092181A1 (fr) * | 2020-10-30 | 2022-05-05 | 東洋紡株式会社 | Anticorps capable de se lier spécifiquement à la protéine de nucléocapside du coronavirus-2 responsable du syndrome respiratoire aigu sévère ou à un fragment de celui-ci, et utilisation dudit anticorps ou dudit fragment |
| JP2022174540A (ja) * | 2021-05-11 | 2022-11-24 | 富士レビオ株式会社 | ＳＡＲＳ－ＣｏＶ－２の免疫学的検出方法および試薬 |
| WO2023277143A1 (fr) * | 2021-06-30 | 2023-01-05 | 株式会社タウンズ | Anticorps dirigé contre la protéine de nucléocapside du sars-cov-2 |
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2022
- 2022-03-04 JP JP2022033238A patent/JP2023128698A/ja active Pending
-
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- 2023-03-03 WO PCT/JP2023/008064 patent/WO2023167317A1/fr unknown
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| WO2021181994A1 (fr) * | 2020-03-10 | 2021-09-16 | デンカ株式会社 | Épitope d'anticorps dirigé contre la protéine structurelle du sras-cov-2, anticorps réagissant avec l'épitope, procédé de détection de sras-cov-2 à l'aide d'anticorps, kit de détection pour anticorps contenant sras-cov-2, procédé de détection d'anticorps anti-sras-cov-2 contenant un polypeptide d'épitope, kit de détection pour un anticorps anti-sras-cov-2 contenant un polypeptide d'épitope, vaccin contre sras-cov-2 contenant un polypeptide d'épitope, et agent thérapeutique pour une infection à sras-cov-2 contenant l'anticorps |
| CN111647053A (zh) * | 2020-04-16 | 2020-09-11 | 军事科学院军事医学研究院生命组学研究所 | 多肽及其在新型冠状病毒检测、抗体或疫苗筛选中的应用 |
| CN113234145A (zh) * | 2020-08-12 | 2021-08-10 | 中山大学附属第五医院 | 特异性结合新型冠状病毒的抗体 |
| WO2022092181A1 (fr) * | 2020-10-30 | 2022-05-05 | 東洋紡株式会社 | Anticorps capable de se lier spécifiquement à la protéine de nucléocapside du coronavirus-2 responsable du syndrome respiratoire aigu sévère ou à un fragment de celui-ci, et utilisation dudit anticorps ou dudit fragment |
| JP2022174540A (ja) * | 2021-05-11 | 2022-11-24 | 富士レビオ株式会社 | ＳＡＲＳ－ＣｏＶ－２の免疫学的検出方法および試薬 |
| WO2023277143A1 (fr) * | 2021-06-30 | 2023-01-05 | 株式会社タウンズ | Anticorps dirigé contre la protéine de nucléocapside du sars-cov-2 |
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| TERRY JAMES S.; ANDERSON LORAN BR.; SCHERMAN MICHAEL S.; MCALISTER CARLEY E.; PERERA RUSHIKA; SCHOUNTZ TONY; GEISS BRIAN J.: "Development of a SARS-CoV-2 nucleocapsid specific monoclonal antibody", VIROLOGY, ELSEVIER, AMSTERDAM, NL, vol. 558, 1 February 2021 (2021-02-01), AMSTERDAM, NL , pages 28 - 37, XP086530250, ISSN: 0042-6822, DOI: 10.1016/j.virol.2021.01.003 * |
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| JP2023128698A (ja) | 2023-09-14 |
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