US 2012/0238037 A1Published application

Immunological assay and immunological assay kit

AssigneeCanon Kabushiki Kaisha

InventorMasaaki Kobayashi

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Abstract

An antibody enabling a rapid measurement of HMGB1, for example a combination of antibodies to HMGB1, is elucidated and an immunological assay and an immunological assay kit are provided. It is intended to provide an immunological assay kit for determining the presence or absence, or the concentration, or both of HMGB1, including a first antibody and a second antibody, wherein the first antibody is immobilized on a carrier and the second antibody is modified with a labeling substance, and the first antibody and the second antibody are any of the following antibody (A), antibody (B), antibody (C), and antibody (D), and a permutation combination of the first antibody and the second antibody is any one of antibody (A) and antibody (B), antibody (A) and antibody (C), antibody (B) and antibody (A), antibody (B) and antibody (C), antibody (B) and antibody (D), and antibody (C) and antibody (B).

Claims3

Dependent claims

An immunological assay for determining the presence or absence, or the concentration, or both of HMGB1 using a first antibody and a second antibody, wherein the first antibody is immobilized on a carrier and the second antibody is modified with a labeling substance, and the first antibody and the second antibody are any of the following antibody (A), antibody (B), antibody (C), and antibody (D), and a permutation combination of the first antibody and the second antibody is any one of antibody (A) and antibody (B), antibody (A) and antibody (C), antibody (B) and antibody (A), antibody (B) and antibody (C), antibody (B) and antibody (D), and antibody (C) and antibody (B):

Antibody (A) is a polyclonal antibody specific for human HMGB1 wherein antibody (A) is produced by and obtained from a rabbit immunized with a protein, as an immunogen, obtained by modifying a peptide represented by
(SEQ ID NO:
2)
GKGDPKKPRGKMSSYA
with KLH (Keyhole Limpet Hemocyanin);
Antibody (B) is a polyclonal antibody specific for human HMGB1 wherein antibody (B) is produced by and obtained from a rabbit immunized with a protein, as an immunogen, represented by
(SEQ ID NO:
3)
MGKGDPKKPRGKMSSYAFFVQTCREEHKKKHPDASVNFSEFSKKCSERW
KTMSAKEKGKFEDMAKADKARYEREMKTYIPPKGETKKKFKDPNAPKRP
PSAFFLFCSEYRPKIKGEHPGLSIGDVAKKLGEMWNNTAADDKQPYEKK
AAKLKEKYEKDIAAYRAKGKPDAAKKGVVKAEKSKKKKEEEEDEEDEED
EEEEEDEEDEDEEEDDDDE;
Antibody (C) is a monoclonal antibody specific for human HMGB1 wherein antibody (C) is produced by and obtained from a mouse immunized with a protein, as an immunogen, obtained by modifying a polypeptide represented by
(SEQ ID NO:
4)
MGKGDPKKPRGKMSSYAFFVQTCREEHKKKHPDASVNFSEFSKKCSERW
KTMSAKEKGKFEDMAKADKARYEREMKTYIPPKGETKKKFK
with a GST (glutathione-S-transferase) tag;
and
Antibody (D) is a monoclonal antibody specific for human HMGB1 wherein antibody (D) is produced by and obtained from a mouse immunized with a protein, as an immunogen, obtained by modifying a protein represented by
(SEQ ID NO:
5)
MGKGDPKKPRGKMSSYAFFVQTCREEHKKKHPDASVNFSEFSKKCSERW
KTMSAKEKGKFEDMAKADKARYEREMKTYIPPKGETKKKFKDPNAPKRP
PSAFFLFCSEYRPKIKGEHPGLSIGDVAKKLGEMWNNTAADDKQPYEKK
AAKLKEKYEKDIAAYRAKGKPDAAKKGVVKAEKSKKKKEEEEDEEDEED
EEEEEDEEDEDEEEDDDDE
with a GST (glutathione-S-transferase) tag.

26 more elements

An immunological assay kit for determining the presence or absence, or the concentration, or both of HMGB1, comprising a first antibody and a second antibody, wherein the first antibody is immobilized on a carrier and the second antibody is modified with a labeling substance, and the first antibody and the second antibody are any of the following antibody (A), antibody (B), antibody (C), and antibody (D), and a permutation combination of the first antibody and the second antibody is any one of antibody (A) and antibody (B), antibody (A) and antibody (C), antibody (B) and antibody (A), antibody (B) and antibody (C), antibody (B) and antibody (D), and antibody (C) and antibody (B):

Antibody (A) is a polyclonal antibody specific for human HMGB1 wherein antibody (A) is produced by and obtained from a rabbit immunized with a protein, as an immunogen, obtained by modifying a peptide represented by
(SEQ ID NO:
2)
GKGDPKKPRGKMSSYA
with KLH (Keyhole Limpet Hemocyanin);
Antibody (B) is a polyclonal antibody specific for human HMGB1 wherein antibody (B) is produced by and obtained from a rabbit immunized with a protein, as an immunogen, represented by
(SEQ ID NO:
3)
MGKGDPKKPRGKMSSYAFFVQTCREEHKKKHPDASVNFSEFSKKCSERW
KTMSAKEKGKFEDMAKADKARYEREMKTYIPPKGETKKKFKDPNAPKRP
PSAFFLFCSEYRPKIKGEHPGLSIGDVAKKLGEMWNNTAADDKQPYEKK
AAKLKEKYEKDIAAYRAKGKPDAAKKGVVKAEKSKKKKEEEEDEEDEED
EEEEEDEEDEDEEEDDDDE;
Antibody (C) is a monoclonal antibody specific for human HMGB1 wherein antibody (C) is produced by and obtained from a mouse immunized with a protein, as an immunogen, obtained by modifying a polypeptide represented by
(SEQ ID NO:
4)
MGKGDPKKPRGKMSSYAFFVQTCREEHKKKHPDASVNFSEFSKKCSERW
KTMSAKEKGKFEDMAKADKARYEREMKTYIPPKGETKKKFK
with a GST (glutathione-S-transferase) tag;
and
Antibody (D) is a monoclonal antibody specific for human HMGB1 wherein antibody (D) is produced by and obtained from a mouse immunized with a protein, as an immunogen, obtained by modifying a protein represented by
(SEQ ID NO:
5)
MGKGDPKKPRGKMSSYAFFVQTCREEHKKKHPDASVNFSEFSKKCSERW
KTMSAKEKGKFEDMAKADKARYEREMKTYIPPKGETKKKFKDPNAPKRP
PSAFFLFCSEYRPKIKGEHPGLSIGDVAKKLGEMWNNTAADDKQPYEKK
AAKLKEKYEKDIAAYRAKGKPDAAKKGVVKAEKSKKKKEEEEDEEDEED
EEEEEDEEDEDEEEDDDDE
with a GST (glutathione-S-transferase) tag.

26 more elements

Dependencies are inferred from the claim text.

Facts

Application: 13/497,795Filed 15 Oct 2010
Priority: JP 2009-24712727 Oct 2009
Published: 20 Sep 2012
Term ends: Not estimatedThe source records don’t support an estimate.

Classification9 codes

G01N 21/6428
Fluorescence; Phosphorescence: measuring fluorescence of fluorescent products of reactions or of fluorochrome labelled reactive substances, e.g.…
C07K 16/18
Immunoglobulins [IG], e.g. monoclonal or polyclonal antibodies: against material from animals or humans
G01N 21/648
Specially adapted constructive features of fluorimeters: using evanescent coupling or surface plasmon coupling for the excitation of fluorescence
G01N 33/6893
Involving proteins, peptides or amino acids: related to diseases not provided for elsewhere