Antigen polypeptide and use thereof

Abstract

To provide a polypeptide suitable for antibody titer measurement.SOLUTION: The present invention relates to any of the polypeptides described in the specification.SELECTED DRAWING: None

Description

Technical Field

[0001] The present disclosure relates to an antigen polypeptide and its uses, etc.

Background Art

[0002] Periodontal disease is an inflammatory disease caused by the infection of periodontal pathogenic bacteria in periodontal tissues. Generally, the diagnosis of periodontal disease is performed by comprehensively considering the results of periodontal pocket examination, palpation and bleeding examination, mobility examination, X-ray examination, etc. However, since these examination methods are based on the experience and skills of dentists, etc., there may be differences in the examination results and diagnosis results, and there is a problem of lacking objectivity. Therefore, the development of a method for objectively diagnosing periodontal disease is desired.

[0003] As a method for objectively diagnosing periodontal disease, for example, a method using the IgG antibody titer in serum against periodontal pathogenic bacteria as an index of the infection degree or severity of periodontal disease has been reported. In the method developed by the present inventors, a polypeptide having a specific primary structure isolated from Porphyromonas gingivalis, a periodontal pathogenic bacterium, or a modified polypeptide thereof (periodontal pathogenic bacterium antigen polypeptide) is used, whereby periodontal diseases of a wide range of patients having various immune types can be examined with high accuracy, and high-speed processing can be performed by an automated device or the like (Patent Document 1).

Prior Art Documents

Patent Documents

[0004]

Patent Document 1

Non-Patent Documents

[0005]

Non-Patent Document 1

Non-Patent Document 2

Summary of the Invention

Problems to be Solved by the Invention

[0006] This disclosure aims to provide a polypeptide suitable for antibody titer measurement.

Means for Solving the Problems

[0007] The inventor has found that a polypeptide having a specific amino acid sequence is suitable for antibody titer measurement and has further improved it.

[0008] This disclosure includes, for example, the subject matters described in the following items. Item 1. Any polypeptide described below. (A-1) An antigen polypeptide consisting of a sequence of 5 or more consecutive amino acids contained in the amino acid sequence shown in SEQ ID NO: 1 (A-2) A polypeptide having an amino acid sequence in which 1 to 3 amino acids are deleted, substituted, or added in the amino acid sequence of the antigen polypeptide of (A-1) and having antigenicity (B-1) An antigen polypeptide consisting of a sequence of 5 or more consecutive amino acids contained in the amino acid sequence shown in SEQ ID NO: 2 (B-2) A polypeptide having an amino acid sequence in which 1 to 3 amino acids are deleted, substituted, or added in the amino acid sequence of the antigen polypeptide of (B-1) and having antigenicity (C-1) An antigen polypeptide consisting of a sequence of 5 or more consecutive amino acids contained in the amino acid sequence shown in SEQ ID NO: 3 (C-2) A polypeptide having an amino acid sequence in which 1 to 3 amino acids are deleted, substituted, or added in the amino acid sequence of the antigen polypeptide of (C-1) and having antigenicity An antigen polypeptide consisting of a sequence of 5 or more consecutive amino acids contained in the amino acid sequence shown in SEQ ID NO: 4 (D-2) A polypeptide having an amino acid sequence in which 1 to 3 amino acids are deleted, substituted, or added in the amino acid sequence of the antigen polypeptide of (D-1), and having antigenicity (E-1) An antigen polypeptide consisting of a sequence of 5 or more consecutive amino acids contained in the amino acid sequence shown in SEQ ID NO: 5 (E-2) A polypeptide having an amino acid sequence in which 1 to 3 amino acids are deleted, substituted, or added in the amino acid sequence of the antigen polypeptide of (E-1), and having antigenicity (F-1) An antigen polypeptide consisting of a sequence of 5 or more consecutive amino acids contained in the amino acid sequence shown in SEQ ID NO: 6 (F-2) A polypeptide having an amino acid sequence in which 1 to 3 amino acids are deleted, substituted, or added in the amino acid sequence of the antigen polypeptide of (F-1), and having antigenicity (G-1) An antigen polypeptide consisting of a sequence of 5 or more consecutive amino acids contained in the amino acid sequence shown in SEQ ID NO: 7 (G-2) A polypeptide having an amino acid sequence in which 1 to 3 amino acids are deleted, substituted, or added in the amino acid sequence of the antigen polypeptide of (G-1), and having antigenicity (H-1) An antigen polypeptide consisting of a sequence of 5 or more consecutive amino acids contained in the amino acid sequence shown in SEQ ID NO: 8 (H-2) A polypeptide having an amino acid sequence in which 1 to 3 amino acids are deleted, substituted, or added in the amino acid sequence of the antigen polypeptide of (H-1), and having antigenicity (I-1) An antigen polypeptide consisting of a sequence of 5 or more consecutive amino acids contained in the amino acid sequence shown in SEQ ID NO: 9 (I-2) A polypeptide having an amino acid sequence in which 1 to 3 amino acids are deleted, substituted, or added in the amino acid sequence of the antigen polypeptide of (I-1), and having antigenicity (J-1) An antigen polypeptide consisting of a sequence of 5 or more consecutive amino acids contained in the amino acid sequence shown in SEQ ID NO: 10 A polypeptide having an amino acid sequence in which 1 to 3 amino acids are deleted, substituted, or added in the amino acid sequence of the antigen polypeptide of (J-2)(J-1) and having antigenicity An antigen polypeptide consisting of a sequence of 5 or more consecutive amino acids contained in the amino acid sequence shown in SEQ ID NO: 11 of (K-1) A polypeptide having an amino acid sequence in which 1 to 3 amino acids are deleted, substituted, or added in the amino acid sequence of the antigen polypeptide of (K-2)(K-1) and having antigenicity An antigen polypeptide consisting of a sequence of 5 or more consecutive amino acids contained in the amino acid sequence shown in SEQ ID NO: 12 of (L-1) A polypeptide having an amino acid sequence in which 1 to 3 amino acids are deleted, substituted, or added in the amino acid sequence of the antigen polypeptide of (L-2)(L-1) and having antigenicity An antigen polypeptide consisting of a sequence of 5 or more consecutive amino acids contained in the amino acid sequence shown in SEQ ID NO: 13 of (M-1) A polypeptide having an amino acid sequence in which 1 to 3 amino acids are deleted, substituted, or added in the amino acid sequence of the antigen polypeptide of (M-2)(M-1) and having antigenicity An antigen polypeptide consisting of a sequence of 5 or more consecutive amino acids contained in the amino acid sequence shown in SEQ ID NO: 14 of (N-1) A polypeptide having an amino acid sequence in which 1 to 3 amino acids are deleted, substituted, or added in the amino acid sequence of the antigen polypeptide of (N-2)(N-1) and having antigenicity Item 2. A polypeptide in which at least two polypeptides selected from the group consisting of the polypeptides of (A-1) to (N-2) described in Item 1 are linked Item 3. A reagent for measuring the antibody titer of periodontal pathogenic bacteria, comprising at least one polypeptide selected from the group consisting of the polypeptides of (A-1) to (N-2) described in Item 1 and / or the polypeptide described in Item 2 Item 4. A kit for measuring the antibody titer of periodontal pathogenic bacteria, comprising the reagent for measuring the antibody titer of periodontal pathogenic bacteria described in Item 3 Item 5. A method for measuring the antibody titer against periodontal pathogenic bacteria in a biological sample, comprising contacting the biological sample with at least one polypeptide selected from the group consisting of the polypeptides (A-1) to (N-2) described in Item 1 and / or the polypeptide described in Item 2. A method for measuring the antibody titer against periodontal pathogenic bacteria in a biological sample.

Effect of the Invention

[0009] A polypeptide suitable for antibody titer measurement is provided.

Mode for Carrying Out the Invention

[0010] Hereinafter, each embodiment included in the present disclosure will be described in more detail. The polypeptide included in the present disclosure includes any of the polypeptides described below. (A-1) An antigen polypeptide consisting of a sequence of 5 or more consecutive amino acids contained in the amino acid sequence shown in SEQ ID NO: 1 (A-2) A polypeptide having an amino acid sequence in which 1 to 3 amino acids are deleted, substituted, or added in the amino acid sequence of the antigen polypeptide of (A-1) and having antigenicity (B-1) An antigen polypeptide consisting of a sequence of 5 or more consecutive amino acids contained in the amino acid sequence shown in SEQ ID NO: 2 (B-2) A polypeptide having an amino acid sequence in which 1 to 3 amino acids are deleted, substituted, or added in the amino acid sequence of the antigen polypeptide of (B-1) and having antigenicity (C-1) An antigen polypeptide consisting of a sequence of 5 or more consecutive amino acids contained in the amino acid sequence shown in SEQ ID NO: 3 (C-2) A polypeptide having an amino acid sequence in which 1 to 3 amino acids are deleted, substituted, or added in the amino acid sequence of the antigen polypeptide of (C-1) and having antigenicity (D-1) An antigen polypeptide consisting of a sequence of 5 or more consecutive amino acids contained in the amino acid sequence shown in SEQ ID NO: 4 A polypeptide having an amino acid sequence in which 1 to 3 amino acids are deleted, substituted, or added in the amino acid sequence of the antigen polypeptide of (D-2)(D-1) and having antigenicity An antigen polypeptide consisting of a sequence of 5 or more consecutive amino acids contained in the amino acid sequence shown in SEQ ID NO: 5 A polypeptide having an amino acid sequence in which 1 to 3 amino acids are deleted, substituted, or added in the amino acid sequence of the antigen polypeptide of (E-2)(E-1) and having antigenicity An antigen polypeptide consisting of a sequence of 5 or more consecutive amino acids contained in the amino acid sequence shown in SEQ ID NO: 6 A polypeptide having an amino acid sequence in which 1 to 3 amino acids are deleted, substituted, or added in the amino acid sequence of the antigen polypeptide of (F-2)(F-1) and having antigenicity An antigen polypeptide consisting of a sequence of 5 or more consecutive amino acids contained in the amino acid sequence shown in SEQ ID NO: 7 A polypeptide having an amino acid sequence in which 1 to 3 amino acids are deleted, substituted, or added in the amino acid sequence of the antigen polypeptide of (G-2)(G-1) and having antigenicity An antigen polypeptide consisting of a sequence of 5 or more consecutive amino acids contained in the amino acid sequence shown in SEQ ID NO: 8 A polypeptide having an amino acid sequence in which 1 to 3 amino acids are deleted, substituted, or added in the amino acid sequence of the antigen polypeptide of (H-2)(H-1) and having antigenicity An antigen polypeptide consisting of a sequence of 5 or more consecutive amino acids contained in the amino acid sequence shown in SEQ ID NO: 9 A polypeptide having an amino acid sequence in which 1 to 3 amino acids are deleted, substituted, or added in the amino acid sequence of the antigen polypeptide of (I-2)(I-1) and having antigenicity An antigen polypeptide consisting of a sequence of 5 or more consecutive amino acids contained in the amino acid sequence shown in SEQ ID NO: 10 A polypeptide having an amino acid sequence in which 1 to 3 amino acids are deleted, substituted, or added in the amino acid sequence of the antigen polypeptide of (J-2)(J-1) and having antigenicity An antigen polypeptide consisting of a sequence of 5 or more consecutive amino acids contained in the amino acid sequence shown in SEQ ID NO: 11 A polypeptide having an amino acid sequence in which 1 to 3 amino acids are deleted, substituted, or added in the amino acid sequence of the antigen polypeptide of (K-2)(K-1) and having antigenicity An antigen polypeptide consisting of a sequence of 5 or more consecutive amino acids contained in the amino acid sequence shown in SEQ ID NO: 12 A polypeptide having an amino acid sequence in which 1 to 3 amino acids are deleted, substituted, or added in the amino acid sequence of the antigen polypeptide of (L-2)(L-1) and having antigenicity An antigen polypeptide consisting of a sequence of 5 or more consecutive amino acids contained in the amino acid sequence shown in SEQ ID NO: 13 A polypeptide having an amino acid sequence in which 1 to 3 amino acids are deleted, substituted, or added in the amino acid sequence of the antigen polypeptide of (M-2)(M-1) and having antigenicity An antigen polypeptide consisting of a sequence of 5 or more consecutive amino acids contained in the amino acid sequence shown in SEQ ID NO: 14 A polypeptide having an amino acid sequence in which 1 to 3 amino acids are deleted, substituted, or added in the amino acid sequence of the antigen polypeptide of (N-2)(N-1) and having antigenicity

[0011] In this specification, each of the above polypeptides may be referred to, for example, as "the polypeptide of (A-1) of the present disclosure", "the polypeptide of (A-2) of the present disclosure", etc. Further, the above polypeptides may be collectively referred to as "the polypeptide of the present disclosure".

[0012] Note that the polypeptides consisting of the amino acid sequences shown in SEQ ID NOS: 1 to 14 are part of the Arg-gingipain RgpA protein (SEQ ID NO: 15) derived from Porphyromonas gingivalis (P. g.).

[0013] The polypeptide of (A-1) of the present disclosure is an antigen polypeptide consisting of a sequence of 5 or more consecutive amino acids included in the amino acid sequence shown in SEQ ID NO: 1. SEQ ID NO: 1 is a sequence consisting of 15 amino acids. Thus, the upper limit of the number of amino acids of the polypeptide of (A-1) is 15. Therefore, the number of amino acids of the polypeptide of (A-1) is 5 to 15. The upper or lower limit of the number of such consecutive amino acids may be, for example, 6, may be 7, may be 8, may be 9, may be 10, may be 11, may be 12, may be 13, may be 14. Specifically, it may be 7 to 15. Among them, 12 to 15 is preferable. Generally, since the number of amino acids constituting an epitope is often 5 to 8, an antigen polypeptide consisting of a sequence of 5 or more consecutive amino acids is expected to be suitably used for antibody titer measurement.

[0014] The polypeptide of (A-2) of the present disclosure is a polypeptide having an amino acid sequence in which 1 to 3 amino acids are deleted, substituted, or added in the amino acid sequence of the antigen polypeptide of (A-1) and having antigenicity. In the amino acid sequence of the antigen polypeptide of (A-1), the number of amino acids deleted, substituted, or added may be, for example, 2.

[0015] Techniques for adding mutations such as deletion, substitution, or addition of amino acids to a specific amino acid sequence are known in the art and can be performed using any method. For example, it can be performed using restriction enzyme treatment, treatment with exonuclease, DNA ligase, etc., site-directed mutagenesis, random mutagenesis, etc.

[0016] The polypeptide of (A-2) of the present disclosure may be, for example, a polypeptide consisting of an amino acid sequence having 85% or more identity with the amino acid sequence of the antigen polypeptide of (A-1). The identity may be 90% or more, 91% or more, 92% or more, 93% or more, 94% or more, 95% or more, 96% or more, 97% or more, 98% or more, or 99% or more.

[0017] The identity of the amino acid sequence can be calculated by using the default parameters in the homology algorithm BLAST (Basic local alignment search tool) of the National Center for Biotechnology Information (NCBI) of the United States, http: / / www.ncbi.nlm.nih.gov / BLAST / .

[0018] The polypeptide of (B-1) of the present disclosure is an antigen polypeptide consisting of a sequence of 5 or more consecutive amino acids contained in the amino acid sequence shown in SEQ ID NO: 2. SEQ ID NO: 2 is a sequence consisting of 15 amino acids, so the upper limit of the number of amino acids of the polypeptide of (B-1) is 15. Therefore, the number of amino acids of the polypeptide of (B-1) is 5 to 15. The upper or lower limit of the number of consecutive amino acids may be, for example, 6, 7, 8, 9, 10, 11, 12, 13, or 14. Specifically, it may be 7 to 15. Among them, 12 to 15 is preferable. Generally, the number of amino acids constituting an epitope is often 5 to 8, so it is expected that an antigen polypeptide consisting of a sequence of 5 or more consecutive amino acids can be suitably used for antibody titer measurement.

[0019] The polypeptide of (B-2) of the present disclosure has an amino acid sequence in which 1 to 3 amino acids are deleted, substituted, or added in the amino acid sequence of the antigen polypeptide of (B-1), and is a polypeptide having antigenicity. In the amino acid sequence of the antigen polypeptide of (B-1), the number of amino acids deleted, substituted, or added may be, for example, 2.

[0020] The polypeptide of (B-2) of the present disclosure may be, for example, a polypeptide consisting of an amino acid sequence showing 85% or more identity with the amino acid sequence of the antigen polypeptide of (B-1). The identity may be 90% or more, 91% or more, 92% or more, 93% or more, 94% or more, 95% or more, 96% or more, 97% or more, 98% or more, or 99% or more.

[0021] The polypeptide of (C-1) of the present disclosure is an antigen polypeptide consisting of a sequence of 5 or more consecutive amino acids contained in the amino acid sequence shown in SEQ ID NO: 3. SEQ ID NO: 3 is a sequence consisting of 12 amino acids, so the upper limit of the number of amino acids of the polypeptide of (C-1) is 12. Therefore, the number of amino acids of the polypeptide of (C-1) is 5 to 12. The upper or lower limit of the number of the consecutive amino acids may be, for example, 6, 7, 8, 9, 10, or 11. Specifically, it may be 7 to 12. Among them, 10 to 12 is preferable. Generally, since the number of amino acids constituting an epitope is often 5 to 8, an antigen polypeptide consisting of a sequence of 5 or more consecutive amino acids is expected to be suitably used for antibody titer measurement.

[0022] The polypeptide of (C-2) of the present disclosure has an amino acid sequence in which 1 to 3 amino acids are deleted, substituted, or added in the amino acid sequence of the antigen polypeptide of (C-1), and is a polypeptide having antigenicity. In the amino acid sequence of the antigen polypeptide of (C-1), the number of amino acids deleted, substituted, or added may be, for example, 2.

[0023] The polypeptide of (C-2) of the present disclosure may be, for example, a polypeptide consisting of an amino acid sequence having 85% or more identity with the amino acid sequence of the antigen polypeptide of (C-1). The identity may be 90% or more, 91% or more, 92% or more, 93% or more, 94% or more, 95% or more, 96% or more, 97% or more, 98% or more, or 99% or more.

[0024] The polypeptide of (D-1) of the present disclosure is an antigen polypeptide consisting of a sequence of 5 or more consecutive amino acids contained in the amino acid sequence shown in SEQ ID NO: 4. SEQ ID NO: 4 is a sequence consisting of 15 amino acids, and thus the upper limit of the number of amino acids of the polypeptide of (D-1) is 15. Therefore, the number of amino acids of the polypeptide of (D-1) is 5 to 15. The upper or lower limit of the number of the consecutive amino acids may be, for example, 6, 7, 8, 9, 10, 11, 12, 13, or 14. Specifically, it may be 7 to 15. Among them, 12 to 15 is preferable. Generally, since the number of amino acids constituting an epitope is often 5 to 8, an antigen polypeptide consisting of a sequence of 5 or more consecutive amino acids is expected to be suitably used for antibody titer measurement.

[0025] The polypeptide of (D-2) of the present disclosure is a polypeptide having an amino acid sequence in which 1 to 3 amino acids are deleted, substituted, or added in the amino acid sequence of the antigen polypeptide of (D-1) and having antigenicity. In the amino acid sequence of the antigen polypeptide of (D-1), the number of amino acids deleted, substituted, or added may be, for example, 2.

[0026] The polypeptide of (D-2) of the present disclosure may be, for example, a polypeptide consisting of an amino acid sequence showing 85% or more identity with the amino acid sequence of the antigen polypeptide of (D-1). The identity may be 90% or more, 91% or more, 92% or more, 93% or more, 94% or more, 95% or more, 96% or more, 97% or more, 98% or more, or 99% or more.

[0027] The polypeptide of (E-1) of the present disclosure is an antigen polypeptide consisting of a sequence of 5 or more consecutive amino acids included in the amino acid sequence shown in SEQ ID NO: 5. SEQ ID NO: 5 is a sequence consisting of 27 amino acids, and thus the upper limit of the number of amino acids of the polypeptide of (E-1) is 27. Therefore, the number of amino acids of the polypeptide of (E-1) is 5 to 27. The upper or lower limit of the number of the consecutive amino acids may be, for example, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, or 26. Specifically, it may be 7 to 27. Among them, 12 to 27 is preferable. Generally, since the number of amino acids constituting an epitope is often 5 to 8, an antigen polypeptide consisting of a sequence of 5 or more consecutive amino acids is expected to be suitably used for antibody titer measurement.

[0028] The polypeptide of (E-2) of the present disclosure is a polypeptide having an amino acid sequence in which 1 to 3 amino acids are deleted, substituted, or added in the amino acid sequence of the antigen polypeptide of (E-1) and having antigenicity. In the amino acid sequence of the antigen polypeptide of (E-1), the number of amino acids deleted, substituted, or added may be, for example, 2.

[0029] The polypeptide of (E-2) of the present disclosure may be, for example, a polypeptide consisting of an amino acid sequence showing 85% or more identity with the amino acid sequence of the antigen polypeptide of (E-1). The identity may be 90% or more, 91% or more, 92% or more, 93% or more, 94% or more, 95% or more, 96% or more, 97% or more, 98% or more, or 99% or more.

[0030] The polypeptide of (F-1) of the present disclosure is an antigen polypeptide consisting of a sequence of 5 or more consecutive amino acids contained in the amino acid sequence shown in SEQ ID NO: 6. SEQ ID NO: 6 is a sequence consisting of 18 amino acids, so the upper limit of the number of amino acids of the polypeptide of (F-1) is 18. Therefore, the number of amino acids of the polypeptide of (F-1) is 5 to 18. The upper or lower limit of the number of the consecutive amino acids may be, for example, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, or 17. Specifically, it may be 7 to 18. Among them, 12 to 18 is preferable. Generally, the number of amino acids constituting an epitope is often 5 to 8, so an antigen polypeptide consisting of a sequence of 5 or more consecutive amino acids is expected to be suitably used for antibody titer measurement.

[0031] The polypeptide of (F-2) of the present disclosure is a polypeptide having an amino acid sequence in which 1 to 3 amino acids are deleted, substituted, or added in the amino acid sequence of the antigen polypeptide of (F-1) and having antigenicity. In the amino acid sequence of the antigen polypeptide of (F-1), the number of amino acids deleted, substituted, or added may be, for example, 2.

[0032] The polypeptide of (F-2) of the present disclosure may be, for example, a polypeptide consisting of an amino acid sequence showing 85% or more identity with the amino acid sequence of the antigen polypeptide of (F-1). The identity may be 90% or more, 91% or more, 92% or more, 93% or more, 94% or more, 95% or more, 96% or more, 97% or more, 98% or more, or 99% or more.

[0033] The polypeptide of (G-1) of the present disclosure is an antigen polypeptide consisting of a sequence of 5 or more consecutive amino acids contained in the amino acid sequence shown in SEQ ID NO: 7. SEQ ID NO: 7 is a sequence consisting of 21 amino acids, so the upper limit of the number of amino acids of the polypeptide of (G-1) is 21. Therefore, the number of amino acids of the polypeptide of (G-1) is 5 to 21. The upper or lower limit of the number of such consecutive amino acids may be, for example, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, or 20. Specifically, it may be 7 to 21. Among them, 12 to 21 is preferable. Generally, since the number of amino acids constituting an epitope is often 5 to 8, it is expected that an antigen polypeptide consisting of a sequence of 5 or more consecutive amino acids can be suitably used for antibody titer measurement.

[0034] The polypeptide of (G-2) of the present disclosure is a polypeptide having an amino acid sequence in which 1 to 3 amino acids are deleted, substituted, or added in the amino acid sequence of the antigen polypeptide of (G-1) and having antigenicity. In the amino acid sequence of the antigen polypeptide of (G-1), the number of amino acids deleted, substituted, or added may be, for example, 2.

[0035] The polypeptide of (G-2) of the present disclosure may be, for example, a polypeptide consisting of an amino acid sequence showing 85% or more identity with the amino acid sequence of the antigen polypeptide of (G-1). The identity may be 90% or more, 91% or more, 92% or more, 93% or more, 94% or more, 95% or more, 96% or more, 97% or more, 98% or more, or 99% or more.

[0036] The polypeptide of (H-1) of the present disclosure is an antigen polypeptide consisting of a sequence of 5 or more consecutive amino acids contained in the amino acid sequence shown in SEQ ID NO: 8. SEQ ID NO: 8 is a sequence consisting of 12 amino acids, so the upper limit of the number of amino acids of the polypeptide of (H-1) is 12. Therefore, the number of amino acids of the polypeptide of (H-1) is 5 to 12. The upper or lower limit of the number of such consecutive amino acids may be, for example, 6, 7, 8, 9, 10, or 11. Specifically, it may be 7 to 12. Among them, 10 to 12 is preferable. Generally, since the number of amino acids constituting an epitope is often 5 to 8, an antigen polypeptide consisting of a sequence of 5 or more consecutive amino acids is expected to be suitably used for antibody titer measurement.

[0037] The polypeptide of (H-2) of the present disclosure is a polypeptide having an amino acid sequence in which 1 to 3 amino acids are deleted, substituted, or added in the amino acid sequence of the antigen polypeptide of (H-1) and having antigenicity. In the amino acid sequence of the antigen polypeptide of (H-1), the number of amino acids deleted, substituted, or added may be, for example, 2.

[0038] The polypeptide of (H-2) of the present disclosure may be, for example, a polypeptide consisting of an amino acid sequence showing 85% or more identity with the amino acid sequence of the antigen polypeptide of (H-1). The identity may be 90% or more, 91% or more, 92% or more, 93% or more, 94% or more, 95% or more, 96% or more, 97% or more, 98% or more, or 99% or more.

[0039] The polypeptide of (I-1) of the present disclosure is an antigen polypeptide consisting of a sequence of 5 or more consecutive amino acids contained in the amino acid sequence shown in SEQ ID NO: 9. SEQ ID NO: 9 is a sequence consisting of 12 amino acids, and thus the upper limit of the number of amino acids of the polypeptide of (I-1) is 12. Therefore, the number of amino acids of the polypeptide of (I-1) is 5 to 12. The upper or lower limit of the number of the consecutive amino acids may be, for example, 6, 7, 8, 9, 10, or 11. Specifically, it may be 7 to 12. Among them, 10 to 12 are preferable. Generally, since the number of amino acids constituting an epitope is often 5 to 8, an antigen polypeptide consisting of a sequence of 5 or more consecutive amino acids is expected to be suitably used for antibody titer measurement.

[0040] The polypeptide of (I-2) of the present disclosure is a polypeptide having an amino acid sequence in which 1 to 3 amino acids are deleted, substituted, or added in the amino acid sequence of the antigen polypeptide of (I-1) and having antigenicity. In the amino acid sequence of the antigen polypeptide of (I-1), the number of amino acids deleted, substituted, or added may be, for example, 2.

[0041] The polypeptide of (I-2) of the present disclosure may be, for example, a polypeptide consisting of an amino acid sequence showing 85% or more identity with the amino acid sequence of the antigen polypeptide of (I-1). The identity may be 90% or more, 91% or more, 92% or more, 93% or more, 94% or more, 95% or more, 96% or more, 97% or more, 98% or more, or 99% or more.

[0042] The polypeptide of (J-1) of the present disclosure is an antigen polypeptide consisting of a sequence of 5 or more consecutive amino acids contained in the amino acid sequence shown in SEQ ID NO: 10. SEQ ID NO: 10 is a sequence consisting of 12 amino acids. Therefore, the upper limit of the number of amino acids of the polypeptide of (J-1) is 12. Accordingly, the number of amino acids of the polypeptide of (J-1) is 5 to 12. The upper or lower limit of the number of the consecutive amino acids may be, for example, 6, 7, 8, 9, 10, or 11. Specifically, it may be 7 to 12. Among them, it is preferably 10 to 12. Generally, the number of amino acids constituting an epitope is often 5 to 8. Therefore, an antigen polypeptide consisting of a sequence of 5 or more consecutive amino acids is expected to be suitably used for antibody titer measurement.

[0043] The polypeptide of (J-2) of the present disclosure is a polypeptide having an amino acid sequence in which 1 to 3 amino acids are deleted, substituted, or added in the amino acid sequence of the antigen polypeptide of (J-1) and having antigenicity. In the amino acid sequence of the antigen polypeptide of (J-1), the number of amino acids deleted, substituted, or added may be, for example, 2.

[0044] The polypeptide of (J-2) of the present disclosure may be, for example, a polypeptide consisting of an amino acid sequence showing 85% or more identity with the amino acid sequence of the antigen polypeptide of (J-1). The identity may be 90% or more, 91% or more, 92% or more, 93% or more, 94% or more, 95% or more, 96% or more, 97% or more, 98% or more, or 99% or more.

[0045] The polypeptide of (K-1) of the present disclosure is an antigen polypeptide consisting of a sequence of 5 or more consecutive amino acids contained in the amino acid sequence shown in SEQ ID NO: 11. SEQ ID NO: 11 is a sequence consisting of 15 amino acids. Therefore, the upper limit of the number of amino acids of the polypeptide of (K-1) is 15. Accordingly, the number of amino acids of the polypeptide of (K-1) is 5 to 15. The upper or lower limit of the number of the consecutive amino acids may be, for example, 6, 7, 8, 9, 10, 11, 12, 13, 14. Specifically, it may be 7 to 15. Among them, it is preferably 12 to 15. Generally, since the number of amino acids constituting an epitope is often 5 to 8, an antigen polypeptide consisting of a sequence of 5 or more consecutive amino acids is expected to be suitably used for antibody titer measurement.

[0046] The polypeptide of (K-2) of the present disclosure is a polypeptide having an amino acid sequence in which 1 to 3 amino acids are deleted, substituted, or added in the amino acid sequence of the antigen polypeptide of (K-1) and having antigenicity. In the amino acid sequence of the antigen polypeptide of (K-1), the number of amino acids deleted, substituted, or added may be, for example, 2.

[0047] The polypeptide of (K-2) of the present disclosure may be, for example, a polypeptide consisting of an amino acid sequence showing 85% or more identity with the amino acid sequence of the antigen polypeptide of (K-1). The identity may be 90% or more, 91% or more, 92% or more, 93% or more, 94% or more, 95% or more, 96% or more, 97% or more, 98% or more, or 99% or more.

[0048] The polypeptide of (L-1) of the present disclosure is an antigen polypeptide consisting of a sequence of 5 or more consecutive amino acids contained in the amino acid sequence shown in SEQ ID NO: 12. SEQ ID NO: 12 is a sequence consisting of 15 amino acids, and thus the upper limit of the number of amino acids of the polypeptide of (L-1) is 15. Therefore, the number of amino acids of the polypeptide of (L-1) is 5 to 15. The upper or lower limit of the number of the consecutive amino acids may be, for example, 6, 7, 8, 9, 10, 11, 12, 13, 14. Specifically, it may be 7 to 15. Among them, it is preferably 12 to 15. Generally, the number of amino acids constituting an epitope is often 5 to 8, and thus an antigen polypeptide consisting of a sequence of 5 or more consecutive amino acids is expected to be suitably used for antibody titer measurement.

[0049] The polypeptide of (L-2) of the present disclosure is a polypeptide having an amino acid sequence in which 1 to 3 amino acids are deleted, substituted, or added in the amino acid sequence of the antigen polypeptide of (L-1) and having antigenicity. In the amino acid sequence of the antigen polypeptide of (L-1), the number of amino acids deleted, substituted, or added may be, for example, 2.

[0050] The polypeptide of (L-2) of the present disclosure may be, for example, a polypeptide consisting of an amino acid sequence showing 85% or more identity with the amino acid sequence of the antigen polypeptide of (L-1). The identity may be 90% or more, 91% or more, 92% or more, 93% or more, 94% or more, 95% or more, 96% or more, 97% or more, 98% or more, or 99% or more.

[0051] The polypeptide of (M-1) of the present disclosure is an antigen polypeptide consisting of a sequence of 5 or more consecutive amino acids contained in the amino acid sequence shown in SEQ ID NO: 13. SEQ ID NO: 13 is a sequence consisting of 15 amino acids, and thus the upper limit of the number of amino acids of the polypeptide of (M-1) is 15. Therefore, the number of amino acids of the polypeptide of (M-1) is 5 to 15. The upper or lower limit of the number of the consecutive amino acids may be, for example, 6, 7, 8, 9, 10, 11, 12, 13, 14, or 15. Specifically, it may be 7 to 15. Among them, 12 to 15 is preferable. Generally, the number of amino acids constituting an epitope is often 5 to 8. Therefore, an antigen polypeptide consisting of a sequence of 5 or more consecutive amino acids is expected to be suitably used for antibody titer measurement.

[0052] The polypeptide of (M-2) of the present disclosure is a polypeptide having an amino acid sequence in which 1 to 3 amino acids are deleted, substituted, or added in the amino acid sequence of the antigen polypeptide of (M-1) and having antigenicity. In the amino acid sequence of the antigen polypeptide of (M-1), the number of amino acids deleted, substituted, or added may be, for example, 2.

[0053] The polypeptide of (M-2) of the present disclosure may be, for example, a polypeptide consisting of an amino acid sequence showing 85% or more identity with the amino acid sequence of the antigen polypeptide of (M-1). The identity may be 90% or more, 91% or more, 92% or more, 93% or more, 94% or more, 95% or more, 96% or more, 97% or more, 98% or more, or 99% or more.

[0054] The (N-1) polypeptide of the present disclosure is an antigen polypeptide consisting of a sequence of 5 or more consecutive amino acids contained in the amino acid sequence shown in SEQ ID NO: 14. SEQ ID NO: 14 is a sequence consisting of 12 amino acids, and thus the upper limit of the number of amino acids of the (N-1) polypeptide is 12. Therefore, the number of amino acids of the (N-1) polypeptide is 5 to 12. The upper or lower limit of the number of the consecutive amino acids may be, for example, 6, 7, 8, 9, 10, or 11. Specifically, it may be 7 to 12. Among them, 10 to 12 are preferable. Generally, the number of amino acids constituting an epitope is often 5 to 8. Therefore, an antigen polypeptide consisting of a sequence of 5 or more consecutive amino acids is expected to be suitably used for antibody titer measurement.

[0055] The (N-2) polypeptide of the present disclosure is a polypeptide having an amino acid sequence in which 1 to 3 amino acids are deleted, substituted, or added in the amino acid sequence of the (N-1) antigen polypeptide and having antigenicity. In the amino acid sequence of the (N-1) antigen polypeptide, the number of amino acids deleted, substituted, or added may be, for example, 2.

[0056] The (N-2) polypeptide of the present disclosure may be, for example, a polypeptide consisting of an amino acid sequence showing 85% or more identity with the amino acid sequence of the (N-1) antigen polypeptide. The identity may be 90% or more, 91% or more, 92% or more, 93% or more, 94% or more, 95% or more, 96% or more, 97% or more, 98% or more, or 99% or more.

[0057] The polypeptide of the present disclosure also includes a polypeptide in which at least two polypeptides selected from the group consisting of the above-described (A-1) to (N-2) polypeptides of the present disclosure are linked.

[0058] At least two polypeptides selected from the group consisting of the polypeptides of (A-1) to (N-2) of the present disclosure may be directly linked or may be linked via a linker sequence or the like.

[0059] The number of polypeptides to be linked is not particularly limited, and may be, for example, 2 to 28. The upper or lower limit of this range may be, for example, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, or 27. More specifically, it may be 2 to 20, may be 2 to 10, or may be 2 to 3.

[0060] The polypeptide of the present disclosure preferably does not contain a polypeptide consisting of the amino acid sequence at positions 696 to 716 of the amino acid sequence shown in SEQ ID NO: 15. The polypeptide of the present disclosure preferably does not contain a polypeptide containing the amino acid sequence at positions 696 to 716 of the amino acid sequence shown in SEQ ID NO: 15.

[0061] As used herein, "having antigenicity" means reacting with an antibody against a periodontal pathogen in an antigen-antibody reaction. Examples of the antibody include immunoglobulin antibodies such as IgG antibody, IgM antibody, and IgA antibody, and any of the immunoglobulin antibodies may react in an antigen-antibody reaction. Among them, IgG antibody is preferred.

[0062] Antigenicity can be evaluated by the ELISA method. More specifically, when a polypeptide for evaluating antigenicity is used as an antigen and a positive serum of a periodontal disease patient containing an antibody against a periodontal pathogen is used as a primary antibody, and these antigen-antibody reactions are detected by the ELISA method, the polypeptide is determined to have antigenicity.

[0063] The polypeptide of the present disclosure can be produced using general protein chemical synthesis methods (e.g., liquid phase method and solid phase method) based on the amino acid sequence information shown in SEQ ID NOs: 1 to 14. Further, the above-described polypeptide can be produced by genetic engineering techniques using a polynucleotide encoding the polypeptide.

[0064] The polypeptide of the present disclosure may further contain a tag. Examples of the tag include peptide tags such as His tag and SBP (Streptavidin Binding Peptide) tag; polypeptide tags such as GST (Glutathione-S-transferase) tag and MBP (Maltose Binding Protein) tag. The polypeptide of the present disclosure may further contain a linker sequence.

[0065] The tag and the linker sequence may be bound to the N-terminal side of the polypeptide of the present disclosure or may be bound to the C-terminal side.

[0066] The method for adding the tag and the linker sequence is not particularly limited, and conventional methods and reaction conditions can be adopted.

[0067] The polypeptide of the present disclosure may further have a labeling substance. Examples of the labeling substance include horseradish peroxidase (HRP), alkaline phosphatase (ALP), β-D-galactosidase, biotin, gold colloid, latex beads, chemiluminescent compounds, and the like.

[0068] The present disclosure also includes a reagent for measuring the periodontal pathogen antibody titer containing the polypeptide of the present disclosure. In this specification, the reagent may be referred to as "the reagent of the present disclosure".

[0069] The reagent of the present disclosure may contain other components in addition to the polypeptide of the present disclosure. Examples of other components include solvents such as physiological saline, PBS, phosphate buffer, Tris buffer, borate buffer, and Good Buffer; protein stabilizers such as those containing casein, skim milk, bovine serum albumin (BSA), gelatin, blood proteins or plant proteins as active ingredients, normal animal blood components such as those of mice, rabbits, goats, fetal calves, etc., and normal human serum components; various preservatives; surfactants; enzyme activators such as salts of metals such as zinc and magnesium, etc.

[0070] The present disclosure also encompasses a periodontal pathogen antibody titer measurement kit containing a reagent for measuring the periodontal pathogen antibody titer containing the polypeptide of the present disclosure. In this specification, the kit may be referred to as "the kit of the present disclosure".

[0071] In addition to the reagent of the present disclosure, the kit may optionally contain other reagents, instruments, etc. For example, it includes blocking solution, washing solution, antibody containing a labeling substance, substrate solution (color development / fluorescence / luminescence), etc. Also, the kit may include members (such as membranes) containing, in addition to the reagent of the present disclosure, blocking solution, washing solution, antibody containing a labeling substance, substrate solution (color development / fluorescence / luminescence), etc.

[0072] The present disclosure also encompasses a method for measuring the antibody titer against periodontal pathogens in a biological sample, which includes contacting the biological sample with the polypeptide of the present disclosure. In this specification, the measurement method may be referred to as "the measurement method of the present disclosure".

[0073] The biological sample is not particularly limited, and examples include samples that may contain antibodies against periodontal pathogens (such as IgG antibody, IgA antibody, IgM antibody, etc.). From the viewpoints of ease of collection and handling, etc., the biological sample is preferably blood, saliva, gingival crevicular fluid, feces, tears, nasal mucus, or amniotic fluid. Also, organs such as the liver can be used as samples that may contain antibodies against periodontal pathogens. When using blood as a biological sample, plasma or serum is more preferred, and serum is even more preferred. When using plasma as a biological sample, it may be plasma separated by filter paper, a plasma separator, or the like. Note that the method, conditions, etc. for storing the biological sample collected from the subject are not particularly limited and can be carried out according to conventional methods.

[0074] The subject from which the biological sample is collected is not particularly limited as long as it is a living body in which periodontal pathogenic bacteria infection can occur, and may be a non-human mammal. There is no particular limitation on humans, and they may be healthy individuals, humans suffering from periodontal disease (periodontal disease patients), or humans suspected of suffering from periodontal disease. Humans having a disease associated with periodontal disease or periodontal pathogenic bacteria (for example, rheumatoid arthritis, heart disease, atherosclerotic disease, diabetes, dementia, non-alcoholic fatty liver (NASH / NAFLD), obesity, premature birth, etc.) or humans suspected of having such a disease may also be used. Examples of non-human mammals include mammals bred as pets, livestock, experimental animals, etc. For example, dogs, cats, monkeys, cows, horses, sheep, goats, pigs, rabbits, mice, rats, camels, llamas, etc. can be mentioned.

[0075] The method for collecting a biological sample from the subject is not particularly limited and may be carried out according to conventional methods. Also, the collected biological sample may be used as it is, or after being freeze-dried and stored, the freeze-dried product may be dissolved in an appropriate solvent described later and used. Alternatively, the collected biological sample may be stored frozen as it is, or after being dissolved in an appropriate solvent described later and stored frozen, it may be thawed and used at the time of use.

[0076] Examples of the solvent for dissolving the biological sample include physiological saline, PBS, phosphate buffer, Tris buffer, borate buffer, Good Buffer, etc. These solvents may contain protein stabilizers such as those containing casein, skim milk, bovine serum albumin (BSA), gelatin, blood proteins or plant proteins as active ingredients, normal animal blood components such as those of mice, rabbits, goats, fetal calves, etc., normal human serum components, etc.; various preservatives; surfactants; enzyme activators such as salts of metals such as zinc and magnesium.

[0077] According to the measurement method of the present disclosure, by contacting the polypeptide of the present disclosure with a biological sample, antibodies against periodontal pathogenic bacteria present in the biological sample (for example, IgG antibody, IgA antibody, IgM antibody, etc.) can be detected by utilizing the antigen-antibody reaction with the polypeptide of the present disclosure.

[0078] The measurement method of the present disclosure may further include a step of detecting an antigen-antibody reaction product (a complex containing an antibody and the polypeptide of the present disclosure) between the antibody against periodontal pathogenic bacteria present in the biological sample generated by contacting the biological sample with the polypeptide of the present disclosure and the polypeptide of the present disclosure.

[0079] The method for detecting the antigen-antibody reaction product is not particularly limited, and conventional methods and reaction conditions can be adopted. For example, a method of contacting the antigen-antibody reaction product with an anti-immunoglobulin antibody (for example, anti-IgG antibody, etc.) containing a labeling substance and detecting the labeling substance, or a method of detecting the labeling substance when the polypeptide of the present disclosure contains a labeling substance, etc. can be mentioned.

[0080] The measurement method of the present disclosure may, if necessary, include, for example, a step of immobilizing the antigen on a solid phase, a washing step, a blocking step, a substrate reaction step, etc. These can adopt conventional methods and reaction conditions.

[0081] For the measurement method of the present disclosure, immunological techniques such as ELISA method, CLEIA method, latex agglutination method, dot blot method, immunochromatography method, etc. are preferably used.

[0082] In addition, as used herein, the term “comprising” includes “consisting essentially of” and “consisting of.” Further, the present disclosure encompasses any combination of the constituent elements described herein.

[0083] Moreover, the various characteristics (properties, structures, functions, etc.) described for each embodiment of the present disclosure above may be combined in any manner in defining the subject matter encompassed by the present disclosure. That is, the present disclosure encompasses all subject matter consisting of any combination of the combinable characteristics described herein.

Example

[0084] The content of the present disclosure will be specifically described using the following experimental examples. However, the present disclosure is not limited thereto in any way. Unless otherwise specified below, the experiments are conducted under atmospheric pressure and normal temperature conditions.

[0085] Epitope mapping was performed on the Arg-gingipain RgpA protein (SEQ ID NO: 15) derived from Porphyromonas gingivalis (P. g.). The epitope mapping was commissioned from KINEXUS. The details are as follows.

[0086] Synthesis of arrays by SPOT On a cellulose membrane, SPOT synthesis of a peptide scan of 12 amino acids with a 3 - amino - acid shift of the RgpA protein (1703aa) was performed. SPOT synthesis of a 10 - copy array was carried out using a MultiPep synthesizer (Intavis). As the cellulose membrane, a commercially available amino - PEG cellulose membrane was used. Peptides were synthesized using an aqueous solution of pre - activated amino acids. Cleavage of the amino - acid protecting groups was performed using an aqueous solution of TFA and scavengers (water, triisopropanol, thioanisole). After cleavage, the membrane containing the array was stored at - 20 °C until probing.

[0087] Probing of the array Probing of the arrays synthesized using serum samples (S004 and S002) from P.g. - infected individuals was performed respectively. Serum samples (S004 and S002) were diluted 1:200 and incubated. Detection of the bound proteins was performed by incubating all arrays with a secondary antibody (HRP - conjugated goat anti - human IgA + IgG + IgM polyclonal antibody) diluted 1:30,000, and then detected using enhanced chemiluminescence (ECL). Chemiluminescence scanning was performed with an exposure time of 50 seconds. During this scanning time, 8 images of the scanned arrays were generated. Immediately after probing, two - step regeneration was performed (without adding β - mercaptoethanol (BME)), and peptide arrays were prepared and stored at - 20 °C until the next probing. Before probing with serum samples, negative controls were examined. As a negative control, one copy (copy 1) of the array was probed with the secondary antibody only. The preceding primary - probing step was mimicked by using T - TBS during the corresponding incubation time. Detection was performed using ECL as described above. Stripping of the membrane was also performed as described above immediately after probing. The washed and dried arrays were stored at - 20 °C in preparation for the next use.

[0088] From the results of epitope mapping, for the amino acid sequences of the identified multiple reactive sites, Affinity (reactivity), functionality (function estimated from the sequence), cross-reactivity (cross-reaction with other species), and commonality (common to S004 and S002) were evaluated based on the following evaluation criteria, and the superiority and inferiority of use as a test antigen were comprehensively determined.

[0089] Affinity (reactivity with serum based on epitope mapping results) ◎: strong 〇: moderate △: weak ×: no signal

[0090] Functionality ◎: Sequence strongly related to the toxicity of gingipain Gingipain is a trypsin-like cysteine protease with cysteine at its active center. The toxicity of RgpA is related to both the catalytic domain involved in enzyme activity and the adhesion / hemagglutinin domain involved in the colonization and infection of P.g. to the host (Non-Patent Documents 1 and 2). In RgpB, which has high homology with RgpA, the amino acids at the active site and substrate binding site have been clarified by three-dimensional structure analysis. In addition, a site with a cell adhesion function related to colonization of the host has been reported. These sequences strongly related to toxicity are present on the surface of the three-dimensional structure for maintaining their function and are sites where antibodies are likely to be formed during P.g. infection, so they are considered effective as test antigens.

[0091] Cross-reactivity (homology search by Blast) ◎: Sequence specific to P.gingivalis 〇: Partially identical to the sequences of P.gulae or P.loveana related to animal periodontal diseases other than P.gingivalis △: Partially identical to the sequences of environmental bacteria or organisms of insects (spiders) other than P.gingivalis ×: Partially identical to the sequences of bacteria or viruses that infect humans other than P.gingivalis

[0092] Commonality 〇: Sequences that react with the sera of both S004 and S002

[0093] The results are shown in Table 1 below (note that Table 1 only shows those with an Affinity of ○ or higher). As described above, the evaluation was performed for every 12 amino acids shifted by 3 amino acids each. For consecutive amino acid sequences that had the same effect, they were summarized as one sequence and described in Table 1.

[0094]

Table 1

[0095] As shown in Table 1, it was confirmed that the polypeptides of SEQ ID NOs: 1 to 14 contain sequences that are excellent for antibody titer measurement.

Claims

1. Any polypeptide described below. (A-1) An antigen polypeptide consisting of a sequence of 5 or more consecutive amino acids contained in the amino acid sequence shown in SEQ ID NO: 1 (A-2) A polypeptide having an amino acid sequence in which 1 to 3 amino acids are deleted, substituted, or added in the amino acid sequence of the antigen polypeptide of (A-1) and having antigenicity (B-1) An antigen polypeptide consisting of a sequence of 5 or more consecutive amino acids contained in the amino acid sequence shown in SEQ ID NO: 2 (B-2) A polypeptide having an amino acid sequence in which 1 to 3 amino acids are deleted, substituted, or added in the amino acid sequence of the antigen polypeptide of (B-1) and having antigenicity (C-1) An antigen polypeptide consisting of a sequence of 5 or more consecutive amino acids contained in the amino acid sequence shown in SEQ ID NO: 3 (C-2) A polypeptide having an amino acid sequence in which 1 to 3 amino acids are deleted, substituted, or added in the amino acid sequence of the antigen polypeptide of (C-1) and having antigenicity (D-1) An antigen polypeptide consisting of a sequence of 5 or more consecutive amino acids contained in the amino acid sequence shown in SEQ ID NO: 4 (D-2) A polypeptide having an amino acid sequence in which 1 to 3 amino acids are deleted, substituted, or added in the amino acid sequence of the antigen polypeptide of (D-1) and having antigenicity (E-1) An antigen polypeptide consisting of a sequence of 5 or more consecutive amino acids contained in the amino acid sequence shown in SEQ ID NO: 5 (E-2) A polypeptide having an amino acid sequence in which 1 to 3 amino acids are deleted, substituted, or added in the amino acid sequence of the antigen polypeptide of (E-1) and having antigenicity (F-1) An antigen polypeptide consisting of a sequence of 5 or more consecutive amino acids contained in the amino acid sequence shown in SEQ ID NO: 6 (F-2) A polypeptide having an amino acid sequence in which 1 to 3 amino acids are deleted, substituted, or added in the amino acid sequence of the antigen polypeptide of (F-1) and having antigenicity (G-1) An antigen polypeptide consisting of a sequence of 5 or more consecutive amino acids contained in the amino acid sequence shown in SEQ ID NO: 7 (G-2) A polypeptide having an amino acid sequence in which 1 to 3 amino acids are deleted, substituted, or added in the amino acid sequence of the antigen polypeptide of (G-1) and having antigenicity (H-1) An antigen polypeptide consisting of a sequence of 5 or more consecutive amino acids contained in the amino acid sequence shown in SEQ ID NO: 8 A polypeptide having an amino acid sequence in which 1 to 3 amino acids are deleted, substituted, or added in the amino acid sequence of the antigen polypeptide of (H-2)(H-1), and having antigenicity An antigen polypeptide consisting of a sequence of 5 or more consecutive amino acids contained in the amino acid sequence shown in SEQ ID NO: 9 A polypeptide having an amino acid sequence in which 1 to 3 amino acids are deleted, substituted, or added in the amino acid sequence of the antigen polypeptide of (I-1), and having antigenicity An antigen polypeptide consisting of a sequence of 5 or more consecutive amino acids contained in the amino acid sequence shown in SEQ ID NO: 10 A polypeptide having an amino acid sequence in which 1 to 3 amino acids are deleted, substituted, or added in the amino acid sequence of the antigen polypeptide of (J-1), and having antigenicity An antigen polypeptide consisting of a sequence of 5 or more consecutive amino acids contained in the amino acid sequence shown in SEQ ID NO: 11 A polypeptide having an amino acid sequence in which 1 to 3 amino acids are deleted, substituted, or added in the amino acid sequence of the antigen polypeptide of (K-1), and having antigenicity An antigen polypeptide consisting of a sequence of 5 or more consecutive amino acids contained in the amino acid sequence shown in SEQ ID NO: 12 A polypeptide having an amino acid sequence in which 1 to 3 amino acids are deleted, substituted, or added in the amino acid sequence of the antigen polypeptide of (L-1), and having antigenicity An antigen polypeptide consisting of a sequence of 5 or more consecutive amino acids contained in the amino acid sequence shown in SEQ ID NO: 13 A polypeptide having an amino acid sequence in which 1 to 3 amino acids are deleted, substituted, or added in the amino acid sequence of the antigen polypeptide of (M-1), and having antigenicity An antigen polypeptide consisting of a sequence of 5 or more consecutive amino acids contained in the amino acid sequence shown in SEQ ID NO: 14 A polypeptide having an amino acid sequence in which 1 to 3 amino acids are deleted, substituted, or added in the amino acid sequence of the antigen polypeptide of (N-1), and having antigenicity

2. A polypeptide in which at least two polypeptides selected from the group consisting of the polypeptides of (A-1) to (N-2) according to claim 1 are linked

3. A reagent for measuring the antibody titer of periodontal pathogenic bacteria, comprising at least one polypeptide selected from the group consisting of the polypeptides of (A-1) to (N-2) according to claim 1 and / or the polypeptide according to claim 2.

4. A kit for measuring the antibody titer of periodontal pathogenic bacteria, comprising the reagent for measuring the antibody titer of periodontal pathogenic bacteria according to claim 3.

5. Contacting a biological sample with at least one polypeptide selected from the group consisting of the polypeptides of (A-1) to (N-2) according to claim 1 and / or the polypeptide according to claim 2, A method for measuring the antibody titer against periodontal pathogenic bacteria in a biological sample.

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