JPWO2009054091A1 - ガストリン放出ペプチド前駆体に対する抗体及びその使用 - Google Patents
ガストリン放出ペプチド前駆体に対する抗体及びその使用 Download PDFInfo
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- JPWO2009054091A1 JPWO2009054091A1 JP2009537894A JP2009537894A JPWO2009054091A1 JP WO2009054091 A1 JPWO2009054091 A1 JP WO2009054091A1 JP 2009537894 A JP2009537894 A JP 2009537894A JP 2009537894 A JP2009537894 A JP 2009537894A JP WO2009054091 A1 JPWO2009054091 A1 JP WO2009054091A1
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- C—CHEMISTRY; METALLURGY
- C07—ORGANIC CHEMISTRY
- C07K—PEPTIDES
- C07K16/00—Immunoglobulins [IGs], e.g. monoclonal or polyclonal antibodies
- C07K16/18—Immunoglobulins [IGs], e.g. monoclonal or polyclonal antibodies against material from animals or humans
- C07K16/26—Immunoglobulins [IGs], e.g. monoclonal or polyclonal antibodies against material from animals or humans against hormones ; against hormone releasing or inhibiting factors
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- 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
- G01N33/574—Immunoassay; Biospecific binding assay; Materials therefor for cancer
- G01N33/57484—Immunoassay; Biospecific binding assay; Materials therefor for cancer involving compounds serving as markers for tumor, cancer, neoplasia, e.g. cellular determinants, receptors, heat shock/stress proteins, A-protein, oligosaccharides, metabolites
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- 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
- G01N33/74—Chemical analysis of biological material, e.g. blood, urine; Testing involving biospecific ligand binding methods; Immunological testing involving hormones or other non-cytokine intercellular protein regulatory factors such as growth factors, including receptors to hormones and growth factors
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- C—CHEMISTRY; METALLURGY
- C07—ORGANIC CHEMISTRY
- C07K—PEPTIDES
- C07K2317/00—Immunoglobulins specific features
- C07K2317/30—Immunoglobulins specific features characterized by aspects of specificity or valency
- C07K2317/34—Identification of a linear epitope shorter than 20 amino acid residues or of a conformational epitope defined by amino acid residues
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- G—PHYSICS
- G01—MEASURING; TESTING
- G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
- G01N2333/00—Assays involving biological materials from specific organisms or of a specific nature
- G01N2333/435—Assays involving biological materials from specific organisms or of a specific nature from animals; from humans
- G01N2333/575—Hormones
- G01N2333/5758—Gastrin releasing peptide
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- G—PHYSICS
- G01—MEASURING; TESTING
- G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
- G01N2800/00—Detection or diagnosis of diseases
- G01N2800/56—Staging of a disease; Further complications associated with the disease
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Abstract
Description
凍結保存した検体におけるProGRP測定値を100%として、4℃で1、4、7日間保存した検体におけるProGRPの測定値を表4に示す。
各モノクローナル抗体0.5mgを1mLの0.1Mリン酸ナトリウム緩衝液(pH8.0)に添加し、ジメチルホルムアミドに溶解したSulfo−NHS−Biotin(PIERCE)をモル比IgG:Biotin=1:20となるように添加し、室温で60分間ゆるやかに攪拌した。1.5Mグリシン溶液(pH8.9)を100μL添加して、室温で10分間ゆるやかに攪拌した。PBSを移動相としてゲルろ過を行い、ビオチン化モノクローナル抗体を得た。
96ウエルマイクロプレートの各ウエルに、各モノクローナル抗体を5μg/mLの濃度で抗体固定液(0.5M NaClを含む0.1M炭酸緩衝液、pH9.6)に溶解した溶液100μLを加え、4℃で一晩インキュベートした。0.15M NaClを含む10mMリン酸緩衝液(pH7.3)で2回洗浄後、ブロッキング液(0.5%カゼインナトリウム、2%スクロースを含む10mMリン酸緩衝液、pH7.1)を350μL加え2時間静置しブロッキングした。ブロッキング液を除去後、反応液(1%BSA、0.05%カゼインナトリウム、1%ポリビニルピロリドン、10mM EDTA−2Na、0.15M NaCl、0.05%Tween20を含む0.1Mリン酸ナトリウム緩衝液、pH7.2)100μLと組換え体ProGRP(0、100、500、2000pg/mL)を50μLずつ各ウエルに加え、37℃で1時間反応させた。洗浄液(0.05%Tween20を含む10mMリン酸緩衝液、pH7.3)で5回洗浄し、ビオチン化した各モノクローナル抗体を1μg/mLの濃度で反応液に溶解させたものを100μL添加して室温で30分間反応させた。反応液で希釈したHRP標識をしたアビジンD(Vector)溶液100μLを添加して、室温で30分間反応させた。洗浄液で5回洗浄し、基質溶液(2mg/mLのオルトフェニレンジアミン、30%過酸化水素水0.9μL/mLを含む0.1Mクエン酸リン酸緩衝液、pH5.0)100μLを加え、20分間インキュベートし、2N硫酸を100μL加えて酵素反応を停止させた。マイクロプレートリーダーで492nm(リファレンス波長600nm)の吸光度を測定した。その反応性を、5段階(高い順から、☆;非常に高い反応性、◎;高い反応性、〇;中程度の反応、△;わずかな反応性、×;反応せず)に分けて表5に示した。
Claims (10)
- 配列番号1に示されるアミノ酸配列の47−68番目のアミノ酸配列によって提示されるエピトープを認識する2種以上の異なる抗体を用いて、ガストリン放出ペプチド前駆体及び/又はその分解物を測定する方法。
- サンドイッチ免疫測定法である、請求項1に記載の方法。
- 配列番号1に示されるアミノ酸配列の47−68番目のアミノ酸配列によって提示されるエピトープを認識するモノクローナル抗体。
- 寄託番号FERM P−21308で寄託されたハイブリドーマGCY9が産生する請求項2に記載のモノクローナル抗体。
- 寄託番号FERM P−21309で寄託されたハイブリドーマGCY17が産生する請求項2に記載のモノクローナル抗体。
- 寄託番号FERM P−21308で寄託されたハイブリドーマGCY9。
- 寄託番号FERM P−21309で寄託されたハイブリドーマGCY17。
- 2種以上の異なる抗体の少なくとも一方が請求項4又は5のいずれかに記載の抗体である、請求項1または2に記載の方法。
- 請求項3〜5のいずれかに記載の抗体を含む、ガストリン放出ペプチド前駆体もしくはその分解物を測定するためのキット。
- 癌の診断又は治療効果のモニタリングを行うためのキットである、請求項9に記載のキット。
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JP2009537894A JP5391071B2 (ja) | 2007-10-26 | 2008-09-29 | ガストリン放出ペプチド前駆体に対する抗体及びその使用 |
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JP2007278843 | 2007-10-26 | ||
JP2007278843 | 2007-10-26 | ||
PCT/JP2008/002716 WO2009054091A1 (ja) | 2007-10-26 | 2008-09-29 | ガストリン放出ペプチド前駆体に対する抗体及びその使用 |
JP2009537894A JP5391071B2 (ja) | 2007-10-26 | 2008-09-29 | ガストリン放出ペプチド前駆体に対する抗体及びその使用 |
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JPWO2009054091A1 true JPWO2009054091A1 (ja) | 2011-03-03 |
JP5391071B2 JP5391071B2 (ja) | 2014-01-15 |
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US (1) | US8609424B2 (ja) |
EP (1) | EP2214016B1 (ja) |
JP (1) | JP5391071B2 (ja) |
KR (1) | KR101525127B1 (ja) |
CN (1) | CN102084249B (ja) |
CA (1) | CA2703787C (ja) |
ES (1) | ES2523588T3 (ja) |
WO (1) | WO2009054091A1 (ja) |
Families Citing this family (7)
Publication number | Priority date | Publication date | Assignee | Title |
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EP3112870B1 (en) * | 2014-02-26 | 2020-05-27 | Konica Minolta, Inc. | Sensor chip for surface plasmon-field enhanced fluorescence spectroscopy comprising different blocking agents |
CN104845975A (zh) * | 2015-05-29 | 2015-08-19 | 郑州大学 | 小细胞肺癌标志物胃泌素释放肽前体多肽片段的dna核酸适体 |
CN108279307B (zh) * | 2018-01-30 | 2021-01-26 | 郑州安图生物工程股份有限公司 | 保护和稳定血清或血浆中胃泌素释放肽前体的采血管 |
CN108982858B (zh) * | 2018-07-05 | 2021-06-15 | 陕西师范大学 | 一种基于单克隆抗体检测人proGRP的双夹心ELISA试剂盒 |
CN111349160B (zh) * | 2018-12-24 | 2021-12-03 | 东莞市朋志生物科技有限公司 | 一种抗人胃泌素释放肽前体的重组抗体 |
CN113583124B (zh) * | 2021-06-16 | 2023-12-15 | 中国辐射防护研究院 | 一种抗前胃泌素释放肽单克隆抗体及其制备方法 |
CN114644710A (zh) * | 2022-03-16 | 2022-06-21 | 中国辐射防护研究院 | 一种抗前胃泌素释放肽单链抗体及其制备方法与用途 |
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US4208479A (en) * | 1977-07-14 | 1980-06-17 | Syva Company | Label modified immunoassays |
CA2089212C (en) * | 1992-06-12 | 2004-01-20 | Ken Yamaguchi | Antibodies to human gastrin-releasing peptide precusor and use thereof |
JP3210994B2 (ja) | 1992-06-12 | 2001-09-25 | 株式会社先端生命科学研究所 | ヒトガストリン放出ペプチド前駆体に対する抗体及びその使用 |
US5468494A (en) * | 1993-11-12 | 1995-11-21 | Aphton Corp. | Immunogenic compositions against human gastrin 17 |
DE69637411T2 (de) * | 1996-06-03 | 2009-01-22 | Advanced Life Science Institute, Inc., Wako | Antikörper gegen den menschlichen gastrinfreisetzenden Peptidvorläufer und deren Verwendung |
EP1191096B1 (en) * | 1999-06-28 | 2011-10-12 | Yusuke Nakamura | Ovel protein and dna thereof |
ATE511647T1 (de) * | 2005-04-13 | 2011-06-15 | Advanced Life Science Inst Inc | Gegen gastrin-releasing-peptide-vorläufer gerichteter antikörper |
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EP2214016A1 (en) | 2010-08-04 |
CA2703787A1 (en) | 2009-04-30 |
CA2703787C (en) | 2016-08-23 |
CN102084249B (zh) | 2014-10-29 |
CN102084249A (zh) | 2011-06-01 |
EP2214016B1 (en) | 2014-09-10 |
ES2523588T3 (es) | 2014-11-27 |
EP2214016A4 (en) | 2011-10-19 |
WO2009054091A1 (ja) | 2009-04-30 |
KR101525127B1 (ko) | 2015-06-03 |
US8609424B2 (en) | 2013-12-17 |
US20100248274A1 (en) | 2010-09-30 |
JP5391071B2 (ja) | 2014-01-15 |
KR20100082764A (ko) | 2010-07-19 |
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