JP7135121B2 - 尿素バイオセンサ及び室温における尿素バイオセンサの安定化 - Google Patents
尿素バイオセンサ及び室温における尿素バイオセンサの安定化 Download PDFInfo
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- JP7135121B2 JP7135121B2 JP2020570928A JP2020570928A JP7135121B2 JP 7135121 B2 JP7135121 B2 JP 7135121B2 JP 2020570928 A JP2020570928 A JP 2020570928A JP 2020570928 A JP2020570928 A JP 2020570928A JP 7135121 B2 JP7135121 B2 JP 7135121B2
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- urea
- membrane
- urease
- ammonium
- polysaccharide
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- QGZKDVFQNNGYKY-UHFFFAOYSA-O Ammonium Chemical compound [NH4+] QGZKDVFQNNGYKY-UHFFFAOYSA-O 0.000 claims description 45
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Images
Classifications
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- G01N27/00—Investigating or analysing materials by the use of electric, electrochemical, or magnetic means
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- G01N27/327—Biochemical electrodes, e.g. electrical or mechanical details for in vitro measurements
- G01N27/3275—Sensing specific biomolecules, e.g. nucleic acid strands, based on an electrode surface reaction
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- G01N27/26—Investigating or analysing materials by the use of electric, electrochemical, or magnetic means by investigating electrochemical variables; by using electrolysis or electrophoresis
- G01N27/28—Electrolytic cell components
- G01N27/30—Electrodes, e.g. test electrodes; Half-cells
- G01N27/327—Biochemical electrodes, e.g. electrical or mechanical details for in vitro measurements
- G01N27/3271—Amperometric enzyme electrodes for analytes in body fluids, e.g. glucose in blood
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Description
本出願は、2019年4月5日に出願され、全ての意図及び目的のために参照によってその全体が本明細書中に援用される米国仮特許出願第62/830,191号に対する優先権及びその利益を主張する。
本発明は、室温での酵素安定性並びに長期の貯蔵寿命及び使用寿命を有する尿素バイオセンサ及びその製造方法に関する。
[0001] 酵素バイオセンサは、血液などの患者体液サンプル中のクレアチニン、クレアチン、グルコース、尿素及び乳酸などの多数の分析物を検出するために使用される。したがって、酵素バイオセンサは、患者の病気のポイントオブケア診断を支援する上で特に重要である。
[0017] 本発明は、室温において安定した尿素バイオセンサ、製造方法、及び安定した尿素バイオセンサを収容するカートリッジに関する。センサ及びバイオセンサという用語は、全体を通して互換的に使用される。
[0031] 以下に記載される本発明は、インビトロ診断、特にポイントオブケア用途のために臨床分析計において有用な尿素バイオセンサの酵素安定性を増強し、貯蔵寿命及び使用寿命を延長するためのデバイス及び関連方法に関する。
実施例1
[0058] 以下に記載される実施例1は、スクロース安定化を有する尿素センサを製造するための方法の一実施形態を提供する。
[0070] 実施例2は、3つの主要因子(それぞれ2つの条件)-(i)センサの作製と、作製したセンサのスクロース溶液への暴露との間の遅延時間(2日又は7日)、(ii)センサがスクロース溶液に曝露される間の浸漬時間(2時間又は16時間)、及びスクロース曝露の完了後の乾燥時間(0.5時間又は3時間)を調査のために用いて行われる多因子実験設計を使用して、周囲温度でスクロース処理が上記の尿素センサの安定化に与える効果を評価するための研究である。尿素センサのサブグループをカートリッジに構築し、使用寿命安定性のためにGEM(登録商標)Premier臨床分析計において尿素濃度を測定した。高濃度の尿素(70mg/dL)を有する水性サンプルをカートリッジ内の尿素センサにおいて毎日試験して、尿素センサ測定の直線性を評価した。
[0074] 実施例3は、3週間の使用寿命安定性及び5ヶ月の貯蔵寿命安定性の研究において、本発明に従って製造された複数の製造バッチから得られる尿素センサの周囲温度における安定性データを提供する。
Claims (27)
- アンモニウム選択高分子膜と、
前記アンモニウム選択高分子膜の外表面上のウレアーゼを備える酵素層と、
前記酵素層の表面に配置され、複合膜の一部を形成する高分子拡散膜と、
(i)前記アンモニウム選択高分子膜上に配置された前記酵素層上に配置された前記高分子拡散膜上に配置された多糖、又は(ii)前記アンモニウム選択高分子膜上に配置された前記酵素層上に配置された前記高分子拡散膜上に配置され、かつ前記酵素層に含まれる多糖であり、前記ウレアーゼが前記アンモニウム選択高分子膜上に前記酵素層を形成する前に前記ウレアーゼに加えられる多糖と、
を備え、
前記多糖が、前記高分子拡散膜と前記アンモニウム選択高分子膜の間の前記ウレアーゼの安定な活性を維持するためにあり、
前記高分子拡散膜が、ポリウレタン、ポリ(テトラフルオロエチレン)アイオノマー、ペルフルオロスルホネートアイオノマー NAFION(登録商標)、ポリ-(2-ヒドロキシメチルメタクリレート)、ポリ塩化ビニル、又は酢酸セルロース、又はそれらの少なくともいずれかの混合物、又はそれらの少なくともいずれかのコポリマーを備える高分子化合物を備える、
尿素バイオセンサ。 - 銀、白金、又は金を備える金属素子と、
前記金属素子と前記アンモニウム選択高分子膜の間の内部電解質溶液と、
をさらに備える、請求項1に記載の尿素バイオセンサ。 - 銀/塩化銀を備える金属素子と、
前記金属素子と前記アンモニウム選択高分子膜の間の内部電解質溶液と、
をさらに備える、請求項1に記載の尿素バイオセンサ。 - 前記高分子拡散膜が、ポリマーマトリックスと、前記ポリマーマトリックス中のアンモニウム選択イオノフォアと、を備える、請求項1に記載の尿素バイオセンサ。
- 前記ポリマーマトリックスが、ポリ塩化ビニル、ポリウレタン、ポリ(テトラフルオロエチレン)、ポリ(メチルメタクリレート)、若しくはシリコーンゴム、又はポリ塩化ビニル、ポリウレタン、ポリ(テトラフルオロエチレン)、ポリ(メチルメタクリレート)、若しくはシリコーンゴムの混合物の一若しくは複数を備える、請求項4に記載の尿素バイオセンサ。
- 前記ポリマーマトリックスがポリ塩化ビニルを備える、請求項4に記載の尿素バイオセンサ。
- 前記アンモニウム選択イオノフォアが、ノナクチン、モナクチン、ジナクチン、トリナクチン、テトラナクチン、ナラシン、ヘキサオキサヘプタシクロトリテトラコンタン、ベンゾクラウンエーテル、若しくは環状デプシペプチド、又はノナクチン、モナクチン、ジナクチン、トリナクチン、テトラナクチン、ナラシン、ヘキサオキサヘプタシクロトリテトラコンタン、ベンゾクラウンエーテル、若しくは環状デプシペプチドの混合物の一若しくは複数を備える、請求項4に記載の尿素バイオセンサ。
- 前記アンモニウム選択イオノフォアがノナクチンを備える、請求項4に記載の尿素バイオセンサ。
- 前記ウレアーゼが架橋される、請求項1に記載の尿素バイオセンサ。
- 前記多糖が、スクロース、トレハロース、ラフィノース、又はラクチトールの一又は複数を備える、請求項1に記載の尿素バイオセンサ。
- 前記多糖が10%のスクロースを備える、請求項1に記載の尿素バイオセンサ。
- 尿素バイオセンサを製造するための方法であって、
溶液中のウレアーゼを電極のアンモニウムイオン選択膜の外表面にキャストして、酵素層を形成することと、
拡散バリアを前記酵素層の表面上に適用することと、
前記拡散バリアを前記酵素層に適用した後、多糖溶液を当該尿素バイオセンサに適用することと、
当該尿素バイオセンサを乾燥させることと、
を含み、
前記拡散バリアが、ポリウレタン、ポリ(テトラフルオロエチレン)アイオノマー、ペルフルオロスルホネートアイオノマー NAFION(登録商標)、ポリ-(2-ヒドロキシメチルメタクリレート)、ポリ塩化ビニル、又は酢酸セルロース、又はそれらの少なくともいずれかの混合物、又はそれらの少なくともいずれかのコポリマーを備える高分子化合物を備える、
方法。 - 前記ウレアーゼが架橋される、請求項12に記載の方法。
- 前記ウレアーゼが、グルタルアルデヒド、1,4-ジイソシアナトブタン、1,2,7,8-ジエポキシオクタン、又は1,2,9,10-ジエポキシデカンの一又は複数を備える化学物質によって架橋される、請求項12に記載の方法。
- 前記電極が、銀、白金、又は金を備える、請求項12に記載の方法。
- 前記電極が、銀/塩化銀電極を備える、請求項12に記載の方法。
- 前記多糖が、スクロース、トレハロース、ラフィノース、又はラクチトールを備える、請求項13に記載の方法。
- 前記多糖を適用することが、前記電極を多糖溶液に少なくとも30分間曝露することを
含む、請求項12に記載の方法。 - 前記尿素バイオセンサが、周囲温度で5ヶ月間の乾燥貯蔵及び21日間の使用後に、安
定した尿素測定性能を維持するよう構成される、請求項12に記載の方法。 - 請求項1に記載の尿素バイオセンサを収容する使い捨てカートリッジ。
- センサのアレイをさらに備え、前記センサが前記尿素バイオセンサを備える、請求項20に記載の使い捨てカートリッジ。
- 前記溶液中のウレアーゼをキャストする前に、多糖溶液が前記溶液中のウレアーゼに添加される、請求項12に記載の方法。
- 前記多糖溶液が10%のスクロースを備える、請求項13に記載の方法。
- 尿素バイオセンサを製造するための方法であって、
溶液中のウレアーゼをアンモニウム選択高分子膜の外表面にキャストして、酵素層を形成することと、
高分子拡散膜を前記酵素層の表面に適用して、複合膜の一部を形成することと、
(i)前記高分子拡散膜が前記酵素層の前記表面に適用された後に、多糖溶液を当該尿素バイオセンサの少なくとも一部に適用すること、又は(ii)前記高分子拡散膜が前記酵素層の前記表面に適用された後、多糖溶液を当該尿素バイオセンサの少なくとも一部に適用し、かつ、前記溶液中のウレアーゼが前記アンモニウム選択高分子膜の前記外表面上にキャストされる前に、多糖溶液を前記溶液中のウレアーゼに加えることと、
当該尿素バイオセンサを乾燥させることと、
を含み、
前記多糖溶液が、前記高分子拡散膜と前記アンモニウム選択高分子膜の間の前記ウレアーゼの安定な活性を維持し、
前記高分子拡散膜が、ポリウレタン、ポリ(テトラフルオロエチレン)アイオノマー、ペルフルオロスルホネートアイオノマー NAFION(登録商標)、ポリ-(2-ヒドロキシメチルメタクリレート)、ポリ塩化ビニル、又は酢酸セルロース、又はそれらの少なくともいずれかの混合物、又はそれらの少なくともいずれかのコポリマーを備える高分子化合物を備える、
方法。 - 前記ウレアーゼが、グルタルアルデヒド、1,4-ジイソシアナトブタン、1,2,7,8-ジエポキシオクタン、又は1,2,9,10-ジエポキシデカンの一又は複数を備える化学物質によって架橋される、請求項24に記載の方法。
- インビトロ診断を実施するための臨床分析計であって、請求項1に記載の尿素バイオセ
ンサを備える、臨床分析計。 - 尿素バイオセンサであって、
アンモニウム選択高分子膜と、
前記アンモニウム選択高分子膜の外表面上のウレアーゼを備える酵素層と、
前記酵素層の表面に適用され、複合膜の一部を形成する高分子拡散膜と、
(i)前記高分子拡散膜が前記酵素層の前記表面に適用された後に当該尿素バイオセンサの少なくとも一部に適用される多糖、又は(ii)前記高分子拡散膜が前記酵素層の前記表面に適用された後に当該尿素バイオセンサの少なくとも一部に適用され、かつ、前記ウレアーゼが前記アンモニウム選択高分子膜上に前記酵素層を形成する前に前記ウレアーゼに加えられる多糖と、
を備え、
前記多糖が、前記高分子拡散膜と前記アンモニウム選択高分子膜の間の前記ウレアーゼの安定な活性を維持し、
当該尿素バイオセンサが、周囲温度での5か月の乾燥貯蔵及び21日の使用後に、安定した尿素測定性能を維持するよう構成されており、
前記高分子拡散膜が、ポリウレタン、ポリ(テトラフルオロエチレン)アイオノマー、ペルフルオロスルホネートアイオノマー NAFION(登録商標)、ポリ-(2-ヒドロキシメチルメタクリレート)、ポリ塩化ビニル、又は酢酸セルロース、又はそれらの少なくともいずれかの混合物、又はそれらの少なくともいずれかのコポリマーを備える高分子化合物を備える、
尿素バイオセンサ。
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