JP2007127634A - 免疫学的検定のための製品およびプロセス - Google Patents
免疫学的検定のための製品およびプロセス Download PDFInfo
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- 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
- G01N33/5302—Apparatus specially adapted for immunological test procedures
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Abstract
【解決手段】膜上の生物学的物質の位置、性質またはその量に関し得る。本発明の方法は、試薬を供給し除去するための圧力支援レジメントを含み、ならびに極めてわずかな量の液体を使用して、検出する膜中に埋もれた物質から汚染物質の洗浄を可能にする。ブロッティング品質を損なうことなく、遮断、洗浄および抗体結合のステップを約30分で終了させることが可能になる。デバイスは、ブロッティング膜の下にある多孔性支持体と、ブロッティング膜の上にある分流器と、所望の領域で液体を収容し、その液体の始動容積をより低くすることを可能にする、分流器上のウェルとを含む、いくつかの層からなる。好ましくは、分流器が、0.22ミクロン膜など、非結合または低結合で親水性の多孔性膜である。
【選択図】図1
Description
a.真空マニホルドと、真空マニホルド上に配置される多孔性支持体と、多孔性支持体上に配置される、検定する1つまたは複数の生物学的存在を含む1つまたは複数の膜と、膜の上部上に配置される分流器と、分流器の上部上に配置される1つまたは複数のウェルとを設けるステップ。
b.1つまたは複数のウェルに1つまたは複数の試薬を加え、試薬を膜中に引き入れるために真空を施すステップ。
c.1つまたは複数のウェルに1つまたは複数の洗浄剤を加え、洗浄剤およびすべての非結合試薬を分流器、膜および多孔性支持体を通して真空マニホルド中に引き入れるために真空を施すステップ。
d.所望または必要に応じてステップbおよびcを1または複数の追加の回数、繰り返すステップ。
a.真空マニホルドと、真空マニホルド上に配置される多孔性支持体と、多孔性支持体上に配置される、検定する1つまたは複数の生物学的存在を含む1つまたは複数の膜と、膜の上部上に配置される分流器と、分流器の上部上に配置される1つまたは複数のウェルとを設けるステップ。
b.1つまたは複数のウェルに1つまたは複数の試薬を加え、試薬を膜中に引き入れるために真空を施すステップ。
c.1つまたは複数のウェルに1つまたは複数の洗浄剤を加え、洗浄剤およびすべての非結合試薬を、分流器、膜および多孔性支持体を通して真空マニホルド中に引き入れるために真空を施すステップ。
d.ステップbおよびcを1または複数の追加の回数、繰り返すステップ。
a.真空マニホルドと、真空マニホルド上に配置される多孔性支持体と、分流器と、分流器の上部上に配置される1つまたは複数のウェルとを設けるステップ。
b.1つまたは複数の生物学的存在を含む1つまたは複数のブロッティング膜を多孔性支持体上に配置するステップ。
c.1つまたは複数のブロッティング膜の上部上に分流器を配置するステップ。
d.分流器のウェルに液体を加え、分流器、1つまたは複数のブロッティング膜および多孔性支持体を通して液体をマニホルド中に引き出すために真空を施すステップ。
a.マニホルドと、マニホルド上に配置される多孔性支持体と、分流器と、分流器の上部上に配置される1つまたは複数のウェルとを設けるステップ。
b.1つまたは複数の生物学的存在を含む1つまたは複数のブロッティング膜を多孔性支持体上に配置するステップ。
c.1つまたは複数のブロッティング膜の上部上に分流器を配置するステップ。
d.分流器のウェルに液体を加え、分流器、1つまたは複数のブロッティング膜および多孔性支持体を通して液体をマニホルド中に移動させるために正圧を分流器に施すステップ。
1.1時間、1%カゼイン/TBS−T中で膜を遮断する(0.1mL/cm2膜)。
2.抗ERK抗体(TBS−T中1%カゼインを用いて1:10,000に希釈)を1時間加える(0.1mL/cm2膜)。
3.TBS−Tで5分間4回洗浄する(1.0mL/cm2膜)。
4.二次抗体(アルカリ性リン酸塩合成抗ウサギIgG;TBS−T中1%カゼインを用いて1:5,000に希釈)を1時間加える(0.1mL/cm2膜)。
5.TBS−Tで5分間4回洗浄する(1.0mL/cm2膜)。
6.Immobilon(登録商標)Western AP試薬を加え、5分間培養する(0.05mL/cm2膜)。
7.X線フィルムに1分間曝し、次いでフィルム現像に進む。
4 多孔性支持体
6 縁部
8 マニホルド
10、34 ブロッティング膜
12、36 分流器
13 正圧マニホルドまたはカバー
14 ウェル
15、18 ポート
16 上部表面
17 チューブ
20 真空源
22 廃物回収デバイス
30 標準真空マニホルド
32 多孔性支持体構造
38 ウェル構造
Claims (15)
- 免疫学的検定を実施するためのデバイスであって、
多孔性支持体と、多孔性支持体の上部上に配置される分流器と、分流器の上部上に取り付けられる1つまたは複数の試薬ウェルとを含む、デバイス。 - 真空支援の免疫学的検定を実施するためのデバイスであって、
真空マニホルドと、多孔性支持体と、多孔性支持体の上部上に配置される分流器と、分流器の上部上に取り付けられる1つまたは複数の試薬ウェルとを含む、デバイス。 - 真空支援の免疫学的検定を実施するためのデバイスであって、
多孔性支持体と、検定する1つまたは複数の生物学的存在を含み、多孔性支持体の上部上に取り付けられる1つまたは複数の膜と、1つまたは複数の膜の上部上の分流器と、分流器の上部上に取り付けられる1つまたは複数の試薬ウェルとを含む、デバイス。 - 圧力支援の免疫学的検定を実施するためのデバイスであって、
回収マニホルドと、多孔性支持体と、検定する1つまたは複数の生物学的存在を含み、多孔性支持体の上部上に位置付けられる1つまたは複数の膜と、1つまたは複数の膜の上部上に配置される分流器と、分流器の上部上に取り付けられる1つまたは複数の試薬ウェルと、1つまたは複数の試薬ウェルの上部上に取り外し可能に密封され、正圧ガス源に接続されるその内部への入口を含む加圧キャップとを含む、デバイス。 - 分流器および1つまたは複数の試薬ウェルが、一体ユニットである、請求項1に記載のデバイス。
- 分流器が、膜であり、
分流器および1つまたは複数の試薬ウェルが、一体ユニットである、請求項1に記載のデバイス。 - 真空支援の免疫学的検定を実施するためのプロセスであって、
a.真空マニホルドと、真空マニホルド上に配置される多孔性支持体と、検定する1つまたは複数の生物学的存在を含み、多孔性支持体上に配置される1つまたは複数の膜と、1つまたは複数の膜の上部上に配置される分流器と、分流器の上部上に配置される1つまたは複数のウェルとを設けるステップと、
b.1つまたは複数のウェルに1つまたは複数の試薬を加え、試薬を1つまたは複数の膜中に引き入れるために真空を施すステップと、
c.1つまたは複数のウェルに1つまたは複数の洗浄剤を加え、洗浄剤およびすべての非結合試薬を、分流器、1つまたは複数の膜および多孔性支持体を通じて真空マニホルド中に引き入れるために真空を施すステップとを含む、プロセス。 - ステップ(b)および(c)が、1または複数回繰り返される、請求項7に記載のプロセス。
- さらに、1つまたは複数の生物学的存在を検出するステップ(d)を含む、請求項7に記載のプロセス。
- さらに、放射性存在および比色分析存在からなる群から選択する検出メカニズムによる、1つまたは複数の生物学的存在を検出するステップ(d)を含む、請求項7に記載のプロセス。
- ステップ(b)の際、試薬が抗体であり、設定時間の間、真空を施すことなく1つまたは複数のブロッティング膜上/中で培養が可能になる、請求項7に記載のプロセス。
- 洗浄液または試薬含有液を、その中の少なくとも1つを検出する1つまたは複数の生物学的存在を含む1つまたは複数のブロッティング膜に通過させるプロセスであって、
a.真空マニホルドと、真空マニホルド上に配置される多孔性支持体と、分流器と、分流器の上部上に配置される1つまたは複数のウェルとを設けるステップと、
b.1つまたは複数の生物学的存在を含む1つまたは複数のブロッティング膜を多孔性支持体上に配置するステップと、
c.1つまたは複数のブロッティング膜の上部上に分流器を配置し、分流器のウェルに液体を加えるステップと、
d.1つまたは複数のブロッティング膜を通じて液体を引き出すために真空を施すステップとを含む、プロセス。 - 洗浄液または試薬含有液を、その中の少なくとも1つを検出する1つまたは複数の生物学的存在を含むブロッティング膜に通過させるプロセスであって、
a.回収マニホルドと、多孔性支持体と、支持体の上部上に配置される分流器と、分流器の上部上に配置される1つまたは複数のウェルと、正圧ガス源に接続されるその内部中への入口を有し、1つまたは複数のウェルの上部上に取り外し可能に取り付けられる加圧キャップとを含むデバイスを設けるステップと、
b.1つまたは複数の生物学的存在を含むブロッティング膜の1つまたは複数の層を多孔性支持体上に配置するステップと、
c.ブロッティング膜の1つまたは複数の層の上部上に分流器を配置し、分流器の1つまたは複数のウェルに液体を加えるステップと、
d.入口からキャップに正圧を加え、ウェルからブロッティング膜の1つまたは複数の層を通してマニホルド中に液体を移動するステップとを含む、プロセス。 - さらに、試薬回収のために、ホルダの下に回収トレイを含む、請求項1に記載のデバイス。
- さらに、試薬回収のために、ホルダの下に回収トレイを含み、
トレイが、2つ以上の別個のサブトレイに分割される、請求項1に記載のデバイス。
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