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US 2025/0258178 A1Published application
Hrg measuring method using anti-hrg monoclonal antibodies
AssigneesShionogi & Co., Ltd., National University Corporation Okayama University
InventorsTakaoki UCHIUMI, Norio OTA, Youko INOUE, Hiroshi NISHI, Kenichi SHIMOSAKO, Masahiro NISHIBORI, Hiroshi MORIMATSU
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Abstract
An object of the present invention is to provide a monoclonal antibody capable of specifically detecting and capturing HRG in a test sample, a set (combination) thereof, and a method for measuring HRG using the same. As a result of intensive studies, the present inventors have found an antibody capable of detecting and capturing HRG in a test sample with good sensitivity and specificity, a set thereof, and a method for measuring HRG using the same, thereby completing the present invention.
Claims19
A monoclonal antibody or antibody fragment thereof against HRG, comprising a heavy chain variable region and a light chain variable region according to any one of the following 1) to 6):
- 1) a heavy chain variable region comprising a CDR1 consisting of an amino acid sequence in which one to two amino acids may be deleted, substituted, inserted and/or added in the amino acid sequence of SEQ ID NO:41,
- a CDR2 consisting of an amino acid sequence in which one to two amino acids may be deleted, substituted, inserted and/or added in the amino acid sequence of SEQ ID NO:42, and
- a CDR3 consisting of an amino acid sequence in which one to two amino acids may be deleted, substituted, inserted and/or added in the amino acid sequence of SEQ ID NO:43,
- and
- a light chain variable region comprising a CDR1 consisting of an amino acid sequence in which one to two amino acids may be deleted, substituted, inserted and/or added in the amino acid sequence of SEQ ID NO:7,
- a CDR2 consisting of an amino acid sequence in which one to two amino acids may be deleted, substituted, inserted and/or added in the amino acid sequence of SEQ ID NO:45, and
- a CDR3 consisting of an amino acid sequence in which one to two amino acids may be deleted, substituted, inserted and/or added in the amino acid sequence of SEQ ID NO:9,
- 2) a heavy chain variable region comprising a CDR1 consisting of an amino acid sequence in which one to two amino acids may be deleted, substituted, inserted and/or added in the amino acid sequence of SEQ ID NO:33,
- a CDR2 consisting of an amino acid sequence in which one to two amino acids may be deleted, substituted, inserted and/or added in the amino acid sequence of SEQ ID NO:34, and
- a CDR3 consisting of an amino acid sequence in which one to two amino acids may be deleted, substituted, inserted and/or added in the amino acid sequence of SEQ ID NO:35,
- and
- a light chain variable region comprising a CDR1 consisting of an amino acid sequence in which one to two amino acids may be deleted, substituted, inserted and/or added in the amino acid sequence of SEQ ID NO:37,
- a CDR2 consisting of an amino acid sequence in which one to two amino acids may be deleted, substituted, inserted and/or added in the amino acid sequence of SEQ ID NO:38, and
- a CDR3 consisting of an amino acid sequence in which one to two amino acids may be deleted, substituted, inserted and/or added in the amino acid sequence of SEQ ID NO:39,
- 3) a heavy chain variable region comprising a CDR1 consisting of an amino acid sequence in which one to two amino acids may be deleted, substituted, inserted and/or added in the amino acid sequence of SEQ ID NO:3,
- a CDR2 consisting of an amino acid sequence in which one to two amino acids may be deleted, substituted, inserted and/or added in the amino acid sequence of SEQ ID NO:4, and
- a CDR3 consisting of an amino acid sequence in which one to two amino acids may be deleted, substituted, inserted and/or added in the amino acid sequence of SEQ ID NO:5,
- and
- a light chain variable region comprising a CDR1 consisting of an amino acid sequence in which one to two amino acids may be deleted, substituted, inserted and/or added in the amino acid sequence of SEQ ID NO:7,
- a CDR2 consisting of an amino acid sequence in which one to two amino acids may be deleted, substituted, inserted and/or added in the amino acid sequence of SEQ ID NO:8, and
- a CDR3 consisting of an amino acid sequence in which one to two amino acids may be deleted, substituted, inserted and/or added in the amino acid sequence of SEQ ID NO:9,
- 4) a heavy chain variable region comprising a CDR1 consisting of an amino acid sequence in which one to two amino acids may be deleted, substituted, inserted and/or added in the amino acid sequence of SEQ ID NO:11,
- a CDR2 consisting of an amino acid sequence in which one to two amino acids may be deleted, substituted, inserted and/or added in the amino acid sequence of SEQ ID NO:12, and
- a CDR3 consisting of an amino acid sequence in which one to two amino acids may be deleted, substituted, inserted and/or added in the amino acid sequence of SEQ ID NO:13,
- and
- a light chain variable region comprising a CDR1 consisting of an amino acid sequence in which one to two amino acids may be deleted, substituted, inserted and/or added in the amino acid sequence of SEQ ID NO:15,
- a CDR2 consisting of an amino acid sequence in which one to two amino acids may be deleted, substituted, inserted and/or added in the amino acid sequence of SEQ ID NO:16, and
- a CDR3 consisting of an amino acid sequence in which one to two amino acids may be deleted, substituted, inserted and/or added in the amino acid sequence of SEQ ID NO:17, and
- 5) a heavy chain variable region comprising a CDR1 consisting of an amino acid sequence in which one to two amino acids may be deleted, substituted, inserted and/or added in the amino acid sequence of SEQ ID NO:21,
- a CDR2 consisting of an amino acid sequence in which one to two amino acids may be deleted, substituted, inserted and/or added in the amino acid sequence of SEQ ID NO:22, and
- a CDR3 consisting of an amino acid sequence in which one to two amino acids may be deleted, substituted, inserted and/or added in the amino acid sequence of SEQ ID NO:23,
- and
- a light chain variable region comprising a CDR1 consisting of an amino acid sequence in which one to two amino acids may be deleted, substituted, inserted and/or added in the amino acid sequence of SEQ ID NO:25,
- a CDR2 consisting of an amino acid sequence in which one to two amino acids may be deleted, substituted, inserted and/or added in the amino acid sequence of SEQ ID NO:26, and
- a CDR3 consisting of an amino acid sequence in which one to two amino acids may be deleted, substituted, inserted and/or added in the amino acid sequence of SEQ ID NO:27,
- 6) a heavy chain variable region comprising a CDR1 consisting of an amino acid sequence in which one to two amino acids may be deleted, substituted, inserted and/or added in the amino acid sequence of SEQ ID NO:11,
- a CDR2 consisting of an amino acid sequence in which one to two amino acids may be deleted, substituted, inserted and/or added in the amino acid sequence of SEQ ID NO:29, and
- a CDR3 consisting of an amino acid sequence in which one to two amino acids may be deleted, substituted, inserted and/or added in the amino acid sequence of SEQ ID NO:13,
- and
- a light chain variable region comprising a CDR1 consisting of an amino acid sequence in which one to two amino acids may be deleted, substituted, inserted and/or added in the amino acid sequence of SEQ ID NO:15,
- a CDR2 consisting of an amino acid sequence in which one to two amino acids may be deleted, substituted, inserted and/or added in the amino acid sequence of SEQ ID NO:31, and
- a CDR3 consisting of an amino acid sequence in which one to two amino acids may be deleted, substituted, inserted and/or added in the amino acid sequence of SEQ ID NO:17.
39 more elements
A set of monoclonal antibodies or antibody fragments thereof against HRG, for use in measurement of HRG in a test sample, which is selected from the following 1) or 2):
- 1) a capture antibody is a monoclonal antibody or antibody fragment thereof against HRG, comprising:
- a heavy chain variable region comprising a CDR1 consisting of an amino acid sequence in which one to two amino acids may be deleted, substituted, inserted and/or added in the amino acid sequence of SEQ ID NO:41,
- a CDR2 consisting of an amino acid sequence in which one to two amino acids may be deleted, substituted, inserted and/or added in the amino acid sequence of SEQ ID NO:42, and
- a CDR3 consisting of an amino acid sequence in which one to two amino acids may be deleted, substituted, inserted and/or added in the amino acid sequence of SEQ ID NO:43,
- and
- a light chain variable region comprising a CDR1 consisting of an amino acid sequence in which one to two amino acids may be deleted, substituted, inserted and/or added in the amino acid sequence of SEQ ID NO:7,
- a CDR2 consisting of an amino acid sequence in which one to two amino acids may be deleted, substituted, inserted and/or added in the amino acid sequence of SEQ ID NO:45, and
- a CDR3 consisting of an amino acid sequence in which one to two amino acids may be deleted, substituted, inserted and/or added in the amino acid sequence of SEQ ID NO:9, and
- a detection antibody is a monoclonal antibody or antibody fragment thereof against HRG, comprising a heavy chain variable region and a light chain variable region according to any one of the following (i) to (iv):
- (i) a heavy chain variable region comprising a CDR1 consisting of an amino acid sequence in which one to two amino acids may be deleted, substituted, inserted and/or added in the amino acid sequence of SEQ ID NO:11,
- a CDR2 consisting of an amino acid sequence in which one to two amino acids may be deleted, substituted, inserted and/or added in the amino acid sequence of SEQ ID NO:12, and
- a CDR3 consisting of an amino acid sequence in which one to two amino acids may be deleted, substituted, inserted and/or added in the amino acid sequence of SEQ ID NO:13,
- and
- a light chain variable region comprising a CDR1 consisting of an amino acid sequence in which one to two amino acids may be deleted, substituted, inserted and/or added in the amino acid sequence of SEQ ID NO:15,
- a CDR2 consisting of an amino acid sequence in which one to two amino acids may be deleted, substituted, inserted and/or added in the amino acid sequence of SEQ ID NO:16, and
- a CDR3 consisting of an amino acid sequence in which one to two amino acids may be deleted, substituted, inserted and/or added in the amino acid sequence of SEQ ID NO:17,
- (ii) a heavy chain variable region comprising a CDR1 consisting of an amino acid sequence in which one to two amino acids may be deleted, substituted, inserted and/or added in the amino acid sequence of SEQ ID NO:21,
- a CDR2 consisting of an amino acid sequence in which one to two amino acids may be deleted, substituted, inserted and/or added in the amino acid sequence of SEQ ID NO:22, and
- a CDR3 consisting of an amino acid sequence in which one to two amino acids may be deleted, substituted, inserted and/or added in the amino acid sequence of SEQ ID NO:23,
- and
- a light chain variable region comprising a CDR1 consisting of an amino acid sequence in which one to two amino acids may be deleted, substituted, inserted and/or added in the amino acid sequence of SEQ ID NO:25,
- a CDR2 consisting of an amino acid sequence in which one to two amino acids may be deleted, substituted, inserted and/or added in the amino acid sequence of SEQ ID NO:26, and
- a CDR3 consisting of an amino acid sequence in which one to two amino acids may be deleted, substituted, inserted and/or added in the amino acid sequence of SEQ ID NO:27,
- (iii) a heavy chain variable region comprising a CDR1 consisting of an amino acid sequence in which one to two amino acids may be deleted, substituted, inserted and/or added in the amino acid sequence of SEQ ID NO:11,
- a CDR2 consisting of an amino acid sequence in which one to two amino acids may be deleted, substituted, inserted and/or added in the amino acid sequence of SEQ ID NO:29, and
- a CDR3 consisting of an amino acid sequence in which one to two amino acids may be deleted, substituted, inserted and/or added in the amino acid sequence of SEQ ID NO:13,
- and
- a light chain variable region comprising a CDR1 consisting of an amino acid sequence in which one to two amino acids may be deleted, substituted, inserted and/or added in the amino acid sequence of SEQ ID NO:15,
- a CDR2 consisting of an amino acid sequence in which one to two amino acids may be deleted, substituted, inserted and/or added in the amino acid sequence of SEQ ID NO:31, and
- a CDR3 consisting of an amino acid sequence in which one to two amino acids may be deleted, substituted, inserted and/or added in the amino acid sequence of SEQ ID NO:17,
- (iv) a monoclonal antibody or antibody fragment thereof against HRG, comprising a heavy chain variable region comprising a CDR1 consisting of an amino acid sequence in which one to two amino acids may be deleted, substituted, inserted and/or added in the amino acid sequence of SEQ ID NO:33,
- a CDR2 consisting of an amino acid sequence in which one to two amino acids may be deleted, substituted, inserted and/or added in the amino acid sequence of SEQ ID NO:34, and
- a CDR3 consisting of an amino acid sequence in which one to two amino acids may be deleted, substituted, inserted and/or added in the amino acid sequence of SEQ ID NO:35,
- and
- a light chain variable region comprising a CDR1 consisting of an amino acid sequence in which one to two amino acids may be deleted, substituted, inserted and/or added in the amino acid sequence of SEQ ID NO:37,
- a CDR2 consisting of an amino acid sequence in which one to two amino acids may be deleted, substituted, inserted and/or added in the amino acid sequence of SEQ ID NO:38, and
- a CDR3 consisting of an amino acid sequence in which one to two amino acids may be deleted, substituted, inserted and/or added in the amino acid sequence of SEQ ID NO:39,
- 2) a capture antibody is a monoclonal antibody or antibody fragment thereof against HRG, comprising:
- a heavy chain variable region comprising a CDR1 consisting of an amino acid sequence in which one to two amino acids may be deleted, substituted, inserted and/or added in the amino acid sequence of SEQ ID NO:3,
- a CDR2 consisting of an amino acid sequence in which one to two amino acids may be deleted, substituted, inserted and/or added in the amino acid sequence of SEQ ID NO:4, and
- a CDR3 consisting of an amino acid sequence in which one to two amino acids may be deleted, substituted, inserted and/or added in the amino acid sequence of SEQ ID NO:5,
- and
- a light chain variable region comprising a CDR1 consisting of an amino acid sequence in which one to two amino acids may be deleted, substituted, inserted and/or added in the amino acid sequence of SEQ ID NO:7,
- a CDR2 consisting of an amino acid sequence in which one to two amino acids may be deleted, substituted, inserted and/or added in the amino acid sequence of SEQ ID NO:8, and
- a CDR3 consisting of an amino acid sequence in which one to two amino acids may be deleted, substituted, inserted and/or added in the amino acid sequence of SEQ ID NO:9, and
- a detection antibody is a monoclonal antibody or antibody fragment thereof against HRG, comprising a heavy chain variable region and a light chain variable region according to any one of the following (i) to (iii):
- (i) a heavy chain variable region comprising a CDR1 consisting of an amino acid sequence in which one to two amino acids may be deleted, substituted, inserted and/or added in the amino acid sequence of SEQ ID NO:11,
- a CDR2 consisting of an amino acid sequence in which one to two amino acids may be deleted, substituted, inserted and/or added in the amino acid sequence of SEQ ID NO:12, and
- a CDR3 consisting of an amino acid sequence in which one to two amino acids may be deleted, substituted, inserted and/or added in the amino acid sequence of SEQ ID NO:13,
- and
- a light chain variable region comprising a CDR1 consisting of an amino acid sequence in which one to two amino acids may be deleted, substituted, inserted and/or added in the amino acid sequence of SEQ ID NO:15,
- a CDR2 consisting of an amino acid sequence in which one to two amino acids may be deleted, substituted, inserted and/or added in the amino acid sequence of SEQ ID NO:16, and
- a CDR3 consisting of an amino acid sequence in which one to two amino acids may be deleted, substituted, inserted and/or added in the amino acid sequence of SEQ ID NO:17,
- (ii) a heavy chain variable region comprising a CDR1 consisting of an amino acid sequence in which one to two amino acids may be deleted, substituted, inserted and/or added in the amino acid sequence of SEQ ID NO:11,
- a CDR2 consisting of an amino acid sequence in which one to two amino acids may be deleted, substituted, inserted and/or added in the amino acid sequence of SEQ ID NO:29, and
- a CDR3 consisting of an amino acid sequence in which one to two amino acids may be deleted, substituted, inserted and/or added in the amino acid sequence of SEQ ID NO:13,
- and
- a light chain variable region comprising a CDR1 consisting of an amino acid sequence in which one to two amino acids may be deleted, substituted, inserted and/or added in the amino acid sequence of SEQ ID NO:15,
- a CDR2 consisting of an amino acid sequence in which one to two amino acids may be deleted, substituted, inserted and/or added in the amino acid sequence of SEQ ID NO:31, and
- a CDR3 consisting of an amino acid sequence in which one to two amino acids may be deleted, substituted, inserted and/or added in the amino acid sequence of SEQ ID NO:17,
- (iii) a monoclonal antibody or antibody fragment thereof against HRG, comprising a heavy chain variable region comprising a CDR1 consisting of an amino acid sequence in which one to two amino acids may be deleted, substituted, inserted and/or added in the amino acid sequence of SEQ ID NO:33,
- a CDR2 consisting of an amino acid sequence in which one to two amino acids may be deleted, substituted, inserted and/or added in the amino acid sequence of SEQ ID NO:34, and
- a CDR3 consisting of an amino acid sequence in which one to two amino acids may be deleted, substituted, inserted and/or added in the amino acid sequence of SEQ ID NO:35,
- and
- a light chain variable region comprising a CDR1 consisting of an amino acid sequence in which one to two amino acids may be deleted, substituted, inserted and/or added in the amino acid sequence of SEQ ID NO:37,
- a CDR2 consisting of an amino acid sequence in which one to two amino acids may be deleted, substituted, inserted and/or added in the amino acid sequence of SEQ ID NO:38, and
- a CDR3 consisting of an amino acid sequence in which one to two amino acids may be deleted, substituted, inserted and/or added in the amino acid sequence of SEQ ID NO:39.
64 more elements
A method for measuring HRG in a test sample using a capture antibody and a detection antibody, the method comprising:
- forming a sandwich immune complex containing a capture antibody, HRG, and a detection antibody,
- wherein the capture antibody is a monoclonal antibody or antibody fragment thereof against HRG, comprising a heavy chain variable region comprising a CDR1 consisting of an amino acid sequence in which one to two amino acids may be deleted, substituted, inserted and/or added in the amino acid sequence of SEQ ID NO:41,
- a CDR2 consisting of an amino acid sequence in which one to two amino acids may be deleted, substituted, inserted and/or added in the amino acid sequence of SEQ ID NO:42, and
- a CDR3 consisting of an amino acid sequence in which one to two amino acids may be deleted, substituted, inserted and/or added in the amino acid sequence of SEQ ID NO:43,
- and
- a light chain variable region comprising a CDR1 consisting of an amino acid sequence in which one to two amino acids may be deleted, substituted, inserted and/or added in the amino acid sequence of SEQ ID NO:7,
- a CDR2 consisting of an amino acid sequence in which one to two amino acids may be deleted, substituted, inserted and/or added in the amino acid sequence of SEQ ID NO:45, and
- a CDR3 consisting of an amino acid sequence in which one to two amino acids may be deleted, substituted, inserted and/or added in the amino acid sequence of SEQ ID NO:9, and
- the detection antibody is a monoclonal antibody or antibody fragment thereof against HRG, comprising a heavy chain variable region comprising a CDR1 consisting of an amino acid sequence in which one to two amino acids may be deleted, substituted, inserted and/or added in the amino acid sequence of SEQ ID NO:33,
- a CDR2 consisting of an amino acid sequence in which one to two amino acids may be deleted, substituted, inserted and/or added in the amino acid sequence of SEQ ID NO:34, and
- a CDR3 consisting of an amino acid sequence in which one to two amino acids may be deleted, substituted, inserted and/or added in the amino acid sequence of SEQ ID NO:35,
- and
- a light chain variable region comprising a CDR1 consisting of an amino acid sequence in which one to two amino acids may be deleted, substituted, inserted and/or added in the amino acid sequence of SEQ ID NO:37,
- a CDR2 consisting of an amino acid sequence in which one to two amino acids may be deleted, substituted, inserted and/or added in the amino acid sequence of SEQ ID NO:38, and
- a CDR3 consisting of an amino acid sequence in which one to two amino acids may be deleted, substituted, inserted and/or added in the amino acid sequence of SEQ ID NO:39.
12 more elements
A method for measuring HRG in a test sample using a set of monoclonal antibodies or antibody fragments thereof against HRG comprising a combination of a capture antibody and a detection antibody, the method comprising:
- (a) reacting a test sample with a capture antibody immobilized on a solid phase in a reaction solution containing an antibody of the same type as the capture antibody or antibody fragment thereof to produce an immune complex containing the capture antibody immobilized on the solid phase and HRG;
- and
- (b) measuring the immune complex produced in the (a) using a detection antibody.
Dependencies are inferred from the claim text.