JP2010520472A - 尿試料中の検体の濃度を正規化するための方法 - Google Patents
尿試料中の検体の濃度を正規化するための方法 Download PDFInfo
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Abstract
以下の工程:
a)少なくとも1つの検体の濃度を測定する工程;
b)少なくとも1つのコラーゲンまたはコラーゲン断片の濃度を測定する工程;
c)前記少なくとも1つのコラーゲンまたはコラーゲン断片の濃度に対して、前記少なくとも1つの検体の濃度を正規化する工程、
を含む、前記方法。
【選択図】図1
Description
a)少なくとも1つの検体の濃度を測定する工程;
b)少なくとも1つのコラーゲンまたはコラーゲン断片の濃度を測定する工程;
c)前記少なくとも1つのコラーゲンまたはコラーゲン断片の濃度に対して、前記少なくとも1つの検体の濃度を正規化する工程、
を含む、前記方法、
によって達成することができる。
−アフィニティクロマトグラフィー、イオン交換クロマトグラフィー、逆相クロマトグラフィー、疎水性相互作用クロマトグラフィー、ゲル濾過、のような、FPLCまたはHPLCを任意に使用するクロマトグラフィー法;
−SELDI法、ClinProt法のような、特定の材料への吸着;
−ゲル電気泳動またはキャピラリー電気泳動のような、電気泳動法、
を使用することができる。
尿試料を、HPLCを用いてダウンストリーム(downstream)ダイオードアレイ検出器(DAD)により検査した。生データを表に示す。ピークは、DADにおける保持時間およびスペクトルによって同定した。これら4つのコラーゲン断片について予め測定していた標準(standard)と測定(measurement)とを比較し、それぞれについて換算因子を算出した。これら因子の平均(この場合は0.80761277)を、他のすべてのシグナルに対する正規化因子(normalizing factor)として使用した。正規化されたシグナルを、最後の欄に記載する。
尿試料を、ソンボーンカードら(Thongboonkerd et al.)、Kidney Int 62: 1461-1469 (2002)に記載されているような2D電気泳動によって、3〜10のpH範囲にわたって、長さ22cmのポリアクリルアミドゲルを用いて分離した。ゲルをクマシー染色によって染色し、免疫学的方法(ウエスタンブロット)を用いて並行して調製したゲルにおいてコラーゲンを検出した。これら5つのたんぱく質について予め測定した標準を使用し、線形回帰(linear regression)を実施した。この較正式により、更に値を換算した。式は、Y = 103.82x − 127449であった。R2値は0.957であった。
尿試料を、質量分析計を組み合わせたキャピラリー電気泳動によって検査した。それにより27個のコラーゲン断片を同定することができ、それをその後予め測定していた標準と比較した。この後、直線は必ずゼロの値を通るという条件による線形回帰を用いて正規化を行った。対応する測定データおよび較正を、以下の表に示す。
Claims (11)
- 尿試料中の少なくとも1つの検体の濃度を正規化するための方法であって、
以下の工程:
a)少なくとも1つの検体の濃度を測定する工程;
b)少なくとも1つのコラーゲンまたはコラーゲン断片の濃度を測定する工程;
c)前記少なくとも1つのコラーゲンまたはコラーゲン断片の濃度に対して、前記少なくとも1つの検体の濃度を正規化する工程、
を含む、前記方法。 - 検体の濃度を測定する前に、尿試料をいくつかの副次標本にさらに分離することを特徴とする、請求項1に記載の方法。
- 前記分離は、
−アフィニティクロマトグラフィー、イオン交換クロマトグラフィー、逆相クロマトグラフィー、疎水性相互作用クロマトグラフィー、ゲル濾過、のような、FPLCまたはHPLCを任意に使用するクロマトグラフィー法;
−SELDI法、ClinProt法のような、特定の材料への吸着;
−ゲル電気泳動またはキャピラリー電気泳動のような、電気泳動法、
により達成されることを特徴とする、請求項2に記載の方法。 - 前記副次標本中の検体および/またはコラーゲンもしくはコラーゲン断片の含有量の測定は、UV/Vis検出器、蛍光検出器、屈折率検出器、ダイオードアレイ検出器、質量分析計、ゲルスキャナー、染色または誘導体化、およびその後の適当な手段による検出、により達成されることを特徴とする、請求項1〜3のいずれか1項に記載の方法。
- 前記少なくとも1つのコラーゲンまたはコラーゲン断片は、その移動または保持時間および任意に更に、マススペクトル、UV/Visスペクトル、蛍光スペクトル、特定のリガンド、例えば、抗体、に対するアフィニティ、のような適当なパラメータによって同定されることを特徴とする、請求項1〜4のいずれか1項に記載の方法。
- 前記少なくとも1つのコラーゲン断片は、配列番号1〜42のペプチドから選択されることを特徴とする、請求項1〜5のいずれか1項に記載の方法。
- 少なくとも2つまたは少なくとも3つ、好ましくは少なくとも5、10、20、30、40または50のコラーゲンまたはコラーゲン断片の濃度が測定され、かつ正規化のために使用されることを特徴とする、請求項1〜6のいずれか1項に記載の方法。
- 前記正規化は、線形回帰により達成されることを特徴とする、請求項1〜7のいずれか1項に記載の方法。
- 尿試料中の検体濃度の正規化のためのコラーゲンまたはコラーゲン断片の使用。
- 前記コラーゲン断片は、配列番号1〜42のペプチドから選択されることを特徴とする、請求項9に記載の使用。
- 少なくとも2つまたは少なくとも3つ、好ましくは少なくとも5、10、20、30、40または50のコラーゲンまたはコラーゲン断片が、正規化のために使用されることを特徴とする、請求項9または10に記載の使用。
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EP07103684 | 2007-03-07 | ||
EP07103684.2 | 2007-03-07 | ||
PCT/EP2008/052730 WO2008107476A1 (de) | 2007-03-07 | 2008-03-06 | Verfahren zur normierung der konzentration von analyten in einer urinprobe |
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JP2010520472A true JP2010520472A (ja) | 2010-06-10 |
JP5351773B2 JP5351773B2 (ja) | 2013-11-27 |
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US (2) | US20100099196A1 (ja) |
EP (1) | EP2118666B1 (ja) |
JP (1) | JP5351773B2 (ja) |
AT (1) | ATE519116T1 (ja) |
AU (1) | AU2008223791B2 (ja) |
BR (1) | BRPI0808703A2 (ja) |
CA (1) | CA2680198C (ja) |
DK (1) | DK2118666T3 (ja) |
WO (1) | WO2008107476A1 (ja) |
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KR102585392B1 (ko) * | 2022-11-03 | 2023-10-06 | 한국기초과학지원연구원 | 콜라겐펩타이드의 분자량분포도 분석 방법 및 시스템 |
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US20060286602A1 (en) * | 2004-05-10 | 2006-12-21 | Harald Mischak | Method and markers for the diagnosis of renal diseases |
EP1972940A1 (de) * | 2007-03-14 | 2008-09-24 | mosaiques diagnostics and therapeutics AG | Verfahren und Marker zur Diagnose von Nierenerkrankungen |
AU2008309605A1 (en) * | 2007-10-09 | 2009-04-16 | Mosaiques Diagnostics And Therapeutics Ag | Polypeptide marker for the diagnosis of prostate cancer |
EP2051078A1 (de) * | 2007-10-19 | 2009-04-22 | mosaiques diagnostics and therapeutics AG | Verfahren und Marker zur Diagnose von Diabetes Mellitus |
WO2009115570A2 (de) * | 2008-03-19 | 2009-09-24 | Mosaiques Diagnostics And Therapeutics Ag | Verfahren und marker zur diagnose von tubulären nierenschäden und -erkrankungen |
US20110214990A1 (en) * | 2008-09-17 | 2011-09-08 | Mosaiques Diagnostics And Therapeutics Ag | Kidney cell carcinoma |
WO2016125271A1 (ja) * | 2015-02-04 | 2016-08-11 | 株式会社島津製作所 | 質量分析方法、クロマトグラフ質量分析装置、及び質量分析用プログラム |
DE102021211272A1 (de) | 2021-10-06 | 2023-04-06 | Ruhr-Universität Bochum, Körperschaft des öffentlichen Rechts | Zusammensetzung enthaltend künstliche Sauerstoffträger zur Verhinderung von Organschäden in Transplantationsorganen |
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KR102585392B1 (ko) * | 2022-11-03 | 2023-10-06 | 한국기초과학지원연구원 | 콜라겐펩타이드의 분자량분포도 분석 방법 및 시스템 |
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Publication number | Publication date |
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US20100099196A1 (en) | 2010-04-22 |
CA2680198C (en) | 2016-01-19 |
WO2008107476A1 (de) | 2008-09-12 |
BRPI0808703A2 (pt) | 2014-09-02 |
AU2008223791A1 (en) | 2008-09-12 |
JP5351773B2 (ja) | 2013-11-27 |
EP2118666A1 (de) | 2009-11-18 |
EP2118666B1 (de) | 2011-08-03 |
CA2680198A1 (en) | 2008-09-12 |
US20150099312A1 (en) | 2015-04-09 |
DK2118666T3 (da) | 2011-11-21 |
AU2008223791B2 (en) | 2013-10-03 |
ATE519116T1 (de) | 2011-08-15 |
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