JP6053781B2 - ラテラルフローアフィニティーアッセイ用の装置及び方法 - Google Patents
ラテラルフローアフィニティーアッセイ用の装置及び方法 Download PDFInfo
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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
- G01N33/543—Immunoassay; Biospecific binding assay; Materials therefor with an insoluble carrier for immobilising immunochemicals
- G01N33/54366—Apparatus specially adapted for solid-phase testing
- G01N33/54386—Analytical elements
- G01N33/54387—Immunochromatographic test strips
- G01N33/54388—Immunochromatographic test strips based on lateral flow
-
- 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
- G01N33/543—Immunoassay; Biospecific binding assay; Materials therefor with an insoluble carrier for immobilising immunochemicals
- G01N33/54366—Apparatus specially adapted for solid-phase testing
- G01N33/54373—Apparatus specially adapted for solid-phase testing involving physiochemical end-point determination, e.g. wave-guides, FETS, gratings
- G01N33/5438—Electrodes
-
- 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
- G01N33/558—Immunoassay; Biospecific binding assay; Materials therefor using diffusion or migration of antigen or antibody
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Description
(i)固相;及び
(ii)検出器
を含み、
固相の表面は、
(a)サンプルがアプライされる第1の位置;及び
(b)第1の位置から離れた第2の位置
を含み、
分析物に結合する、検出可能な分子種を放出できる第1の分子が、固相上に沈着しているか、固相へのアプライ前にサンプルに添加され、
分析物に結合する第2の分子が、第2の位置に固定化されており、
酵素が、第2の位置で固定化されて固定化分子と共局在しており、
検出器は、第2の位置の固定化分子に近接して配置されている。
a)検出可能な分子種を放出できる第1の可溶性標識結合分子及び第2の可溶性標識結合分子をサンプルに溶解して複合体を形成させるステップ;
b)ステップa)で形成された複合体を固相にアプライするステップであって、
固相の表面が、
i)複合体がアプライされる第1の位置;及び
ii)第1の位置から離れた第2の位置
を含み、第2の標識結合分子に結合する分子が第2の位置に固定化されており、酵素が、第2の位置で固定化されて固定化分子と共局在しており、検出器が第2の位置の固定化分子に近接して配置されている、
ステップ;及び
c)検出可能な分子種を検出器で検出するステップであって、
前記分子種の検出が、サンプルの標的分子含有量に比例する、
ステップ。
。
(i)固相;及び
(ii)検出器
を含む、サンプル中の分析物を定量分析するための装置であって、
固相の表面が、
(a)サンプルがアプライされる第1の位置;及び
(b)第1の位置から離れた第2の位置
を含み、
分析物に結合する、検出可能な分子種を放出できる第1の分子が、固相に沈着しているか、固相へのアプライ前にサンプルに添加され、
分析物に結合する第2の分子が、第2の位置に固定化されており、
酵素が、固定化されて第2の位置の固定化されている分子と共局在しており、
検出器が、第2の位置の固定化分子に近接して配置されている、
装置を提供する。
a)検出可能な分子種を放出できる第1の可溶性標識結合分子及び第2の可溶性標識結合分子をサンプルに溶解させて複合体を形成させるステップ;
b)ステップa)で形成された複合体を固相にアプライするステップであって、固相の表面が、
i)複合体がアプライされる第1の位置;及び
ii)第1の位置から離れた第2の位置、
を含み、
第2の標識結合分子に結合する分子が、第2の位置に固定化されており、酵素が第2
の位置で固定化されて固定化分子と共局在しており、検出器が、第2の位置の固定化分子に近接して配置されている、
ステップ;及び
c)検出可能な分子種を検出器で検出するステップであって、
前記分子種の検出が、サンプル中の標的分子の含有量に比例する、ステップ。
用される具体的な酵素に応じて、より酸性側の環境への変化(すなわち、pH低下)から、よりアルカリ性側の環境への変化(すなわち、pH上昇)であり得る。
Claims (16)
- (i)固相;及び
(ii)検出器
を含む、サンプル中の抗原を定量分析するための装置であって、
前記固相の表面が、
(a)サンプルがアプライされる第1の位置;及び
(b)前記第1の位置から離れた第2の位置
を含み、
前記抗原に結合する、封入レドックスプローブを放出できる第1の抗体が、前記第1の位置に沈着されているか、前記固相へのアプライ前にサンプルに添加され、
前記抗原に結合する第2の抗体が、前記第2の位置に固定化されており、
酵素が、前記第2の位置で固定化されて前記固定化抗体と共局在しており、
前記検出器が、前記第2の位置の前記固定化抗体に近接して配置されている、装置であって、
前記酵素の基質が、サンプル中に通常存在するものであるか、前記固相へのアプライ前にサンプルに添加されるか、サンプルアプライ前に前記固相上の前記第1の位置に沈着され、
前記酵素と前記基質との相互作用により、前記第1の抗体から前記レドックスプローブが放出されるpH変化が生じる、装置。 - 前記検出器による前記レドックスプローブの検出を前記サンプル中の抗原濃度の値へと変換するための手段を更に含む、請求項1に記載の装置。
- 前記検出器が電極である、請求項1又は2に記載の装置。
- 前記酵素がウレアーゼであり、前記基質が尿素である、請求項1〜3のいずれか一項に記載の装置。
- 前記固相がラテラルフローメンブレンである、請求項1〜4のいずれか一項に記載の装置。
- 基質含有サンプル中の標的抗原を定量的に検出する方法であって、
a)封入レドックスプローブを放出できる第1の可溶性標識抗体及び第2の可溶性標識抗体をサンプルに溶解させて免疫複合体を形成させるステップ;
b)ステップa)で形成された前記免疫複合体を固相にアプライするステップであって、表面が、
i)前記免疫複合体がアプライされる第1の位置、及び
ii)前記第1の位置から離れた第2の位置
を含み、
前記第2の抗体に結合する分子が、前記第2の位置に固定化されており、酵素が、前記第2の位置で固定化されて前記固定化分子と共局在しており、検出器が、前記第2の位置の前記固定化分子に近接して配置されている、
ステップ;及び
c)前記免疫複合体が前記第2の位置に輸送された時に前記検出器で前記レドックスプローブを検出するステップであって、前記レドックスプローブの検出が前記サンプルの前記標的抗原含有量に比例する、ステップ
を含み、
前記酵素と前記基質との相互作用により、前記第1の抗体から前記レドックスプローブが放出されるpH変化が生じる、方法。 - 前記第2の抗体がビオチンで標識されており、それに結合する、前記第2の位置に固定化されている分子がアビジンである、請求項6に記載の方法。
- ステップ(c)が、局所的pH変化及びそれによる生じる前記レドックスプローブの放出による電流又は通過する電荷の変化を前記検出器で検出することを含む、請求項6又は7に記載の方法。
- 前記レドックスプローブの酸化又は還元による前記電流が前記サンプル中の抗原含有量に比例する、請求項8に記載の方法。
- 前記検出器が電極である、請求項6〜9のいずれか一項に記載の方法。
- 前記電極がスクリーン印刷電極である、請求項10に記載の方法。
- 前記レドックスプローブがポリマー中に封入されている、請求項6〜11のいずれか一項に記載の方法。
- 前記ポリマーがアルカリ性条件下で溶解する腸溶性コーティングである、請求項12に記載の方法。
- 前記ポリマーが酸性又はアルカリ性条件下で膨潤する、請求項12に記載の方法。
- 前記サンプルが尿素含有サンプルである、請求項6〜14のいずれか一項に記載の方法。
- 前記酵素がウレアーゼである、請求項6〜15のいずれか一項に記載の方法。
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PCT/GB2012/051733 WO2013011323A2 (en) | 2011-07-19 | 2012-07-19 | Apparatus and method for affinity assays |
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US9678070B2 (en) * | 2015-04-29 | 2017-06-13 | Church & Dwight Co., Inc. | Method and apparatus for electrochemical detection |
US20200333311A1 (en) | 2018-01-05 | 2020-10-22 | Simpore Inc. | Sample preparation and flow-through sensors using functionalized silicon nanomembranes |
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EP2734842B1 (en) | 2016-01-20 |
WO2013011323A3 (en) | 2013-03-07 |
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