JP2005300552A - イムノアッセイ用試薬の緩衝剤組成物 - Google Patents
イムノアッセイ用試薬の緩衝剤組成物 Download PDFInfo
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- JP2005300552A JP2005300552A JP2005139663A JP2005139663A JP2005300552A JP 2005300552 A JP2005300552 A JP 2005300552A JP 2005139663 A JP2005139663 A JP 2005139663A JP 2005139663 A JP2005139663 A JP 2005139663A JP 2005300552 A JP2005300552 A JP 2005300552A
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Abstract
【解決手段】免疫学的手順を行うときの使用に適する水性組成物。該組成物は、少なくとも1種の生物学的緩衝剤、ジチオトレイトール(あるいは、「DTT」と言う)、およびエチレングリコールを含む。組成物はまた、少なくとも1種の生物学的洗剤、少なくとも1種の正および負の対イオン源(例えば、塩)、ならびに少なくとも1種の粘度調節剤(例えば、糖)も含み得る。緩衝剤はまた、少なくとも1種の保存剤(アジ化ナトリウムなど)を含み得る。組成物のpHは、好ましくは、約 6.4〜7.2 の範囲である。該組成物を含むキットも開示する。
【選択図】なし
Description
本発明は、少なくとも1種の生物学的緩衝剤、ジチオトレイトール(あるいは、「DTT」とも言う)およびエチレングリコールを含む、緩衝剤としての使用に適する水性組成物を提供する。該組成物の媒体は水である。組成物はまた、少なくとも1種の生物学的洗剤、少なくとも1種の正および負の対イオン源(例えば、塩)、ならびに少なくとも1種の粘度調節剤(例えば、糖)を含むこともできる。緩衝剤はまた、少なくとも1種の保存剤(例えば、アジ化ナトリウム)を含むことができる。本発明の組成物のpHは、約6.4〜約7.2、好ましくは約6.4〜約6.8、より好ましくは約6.5〜約6.7の範囲であり得る。最も好ましくは、組成物のpHは約6.6である。
本発明は、熱ストレス条件下での抗原および抗体の安定性の改善に特に有用な水性組成物に関する。組成物は、次の成分を含む。すなわち、少なくとも1種の生物学的緩衝剤、ジチオトレイトール〔HSCH2(CHOH)2CH2SH〕およびエチレングリコール〔HOCH2CH2OH〕である。組成物の媒体は水である。
ベンゼン−1,2,4,5−テトラカルボン酸(ピロメリット酸);
ベンゼン−1,2,3−トリカルボン酸(ヘミメリット酸);
ジメチルマロン酸;
ヒスチジン;
ヒドロキシルアミン;
炭酸(H2CO3+CO2);
マロン酸;
2−(N−モルホリノ)−エタンスルホン酸「MES」;
グリセロリン酸;
プロパン−1,2,3−トリカルボン酸(トリカルバリル酸);
ベンゼンペンタカルボン酸;
マレイン酸;
2,2−ジメチルコハク酸;
エチレンジアミン四酢酸「EDTA」;
3,3−ジメチルグルタル酸;
ビス(2−ヒドロキシエチル)イミノ−トリス(ヒドロキシメチル)メタン「BIS−TRIS」;
ベンゼンヘキサカルボン酸(メリット酸);
N−(2−アセタミド)イミノ−二酢酸「ADA」;
ブタン−1,2,3,4−テトラカルボン酸;
ピロリン酸;
1,1−シクロペンタン二酢酸(3,3−テトラメチレン−グルタル酸);
1,4−ピペラジンビス−(エタンスルホン酸)「PIPES」;
N−(2−アセタミド)−2−アミノエタンスルホン酸「ACES」;
1,1−シクロヘキサン二酢酸;
3,6−エンドメチレン−1,2,3,6−テトラヒドロフタル酸「EMTA」(「ENDCA」);
イミダゾール;
塩化2−(アミノエチル)トリメチルアンモニウム「CHOLAMINE」;
N,N−ビス(2−ヒドロキシエチル)−2−アミノエタンスルホン酸「BES」;
2−メチルプロパン−1,2,3−トリスカルボン酸(β−メチルトリカルバリル酸);
2−(N−モルホリノ)プロパン−スルホン酸「MOPS」;
リン酸;
N−トリス(ヒドロキシメチル)メチル−2−アミノエタンスルホン酸「TES」;および
N−2−ヒドロキシエチルピペラジン−N’−2−エタンスルホン酸「HEPES」
上記リストにおいて、括弧内の化合物名は、括弧の化合物名の前にある化合物の一般名である。カギ括弧の頭文字語は、カギ括弧の頭文字語の前にある化合物を指示するとき、一般に使用される化合物名である。
本実施例は、生物学的緩衝剤、ジチオトレイトールおよびエチレングリコールを含む組成物の、熱ストレス条件下でのC型肝炎ウイルス(HCV)抗原の安定性に対する効果を示す。
31℃
37℃
グリセロールを含む溶液は、アッセイでのシグナル発生を激しく減少させた。従って、この溶液は、4日以上は評価を行わなかった。ポリ酢酸ビニルを含む溶液も、アッセイでのシグナル発生を減少させた。従って、この溶液は、4日以上の評価を行わなかった。グリコールおよびポリエチレングリコールを含む溶液ならびに対照は、その各々の温度で維持され、12日目の試験を行った。
大腸菌で発現される組換えHCVタンパク質HC43は、推定上のHCVコア構造タンパク質およびHCV非構造タンパク質NS3の配列を含む。HC43は、HCVポリタンパク質のアミノ酸 1〜150 および1192〜1457から成る融合タンパク質である。HCVポリタンパク質のアミノ酸1192〜1457は、33c抗原を構成する。
サッカロミセス・セレビジエ(Saccharomyces cerevisiae)(酵母)で発現される組換えHCVタンパク質c100−3は、推定上のHCV非構造タンパク質NS3およびNS4の配列を含む。c100−3は、ヒトスーパーオキシドジスムターゼ(SOD)の 154個のアミノ酸、5個のリンカーアミノ酸、HCVポリタンパク質のアミノ酸1569〜1931およびカルボキシ末端の付加的な5個のリンカーアミノ酸のキメラ融合領域である。
酵母で発現される組換えHCVタンパク質c200は、HCVゲノムのNS3およびNS4領域をコードすると思われるHCVアミノ酸1192〜1931を含む。c200は、スーパーオキシドジスムターゼ(SOD)の 154個のアミノ酸を有するキメラ融合タンパク質である。
酵母で発現される組換えHCVタンパク質NS5は、推定上のHCV非構造タンパク質NS5の配列を含む。NS5は、SODの154個のアミノ酸およびHCVポリタンパク質のアミノ酸2054〜2995のキメラ融合領域である。
Claims (23)
- 熱ストレス条件下における、ウイルスゲノムのNS3領域のC型肝炎ウイルス抗原に係る免疫応答性の安定化に適当な水性組成物であって、
該組成物は、pHを7.2以下のレベルで維持するのに十分な濃度の少なくとも一種の生物学的緩衝剤;
熱ストレス条件下における、ウイルスゲノムのNS3領域の該C型肝炎ウイルス抗原に係るスルフヒドリル基を保護するのに十分な濃度のジチオトレイトール;
4%〜8%(%は、g/100mlを意味する)の範囲の濃度で存在するエチレングリコールを含み、組成物のpHが7.2以下であることを特徴とする該組成物。 - 少なくとも1種の生物学的緩衝剤が、10mM〜500mMの範囲の濃度で存在する、請求項1に記載の組成物。
- 少なくとも1種の生物学的緩衝剤が、
ベンゼン−1,2,4,5−テトラカルボン酸(ピロメリット酸);
ベンゼン−1,2,3−トリカルボン酸(ヘミメリット酸);
ジメチルマロン酸;
ヒスチジン;
ヒドロキシルアミン;
炭酸(H2CO3+CO2);
マロン酸;
2−(N−モルホリノ)−エタンスルホン酸「MES」;
グリセロリン酸;
プロパン−1,2,3−トリカルボン酸(トリカルバリル酸);
ベンゼンペンタカルボン酸;
マレイン酸;
2,2−ジメチルコハク酸;
エチレンジアミン四酢酸「EDTA」;
3,3−ジメチルグルタル酸;
ビス(2−ヒドロキシエチル)イミノ−トリス(ヒドロキシメチル)メタン「BIS−TRIS」;
ベンゼンヘキサカルボン酸(メリット酸);
N−(2−アセタミド)イミノ−二酢酸「ADA」;
ブタン−1,2,3,4−テトラカルボン酸;
ピロリン酸;
1,1−シクロペンタン二酢酸(3,3−テトラメチレン−グルタル酸);
1,4−ピペラジンビス−(エタンスルホン酸)「PIPES」;
N−(2−アセタミド)−2−アミノエタンスルホン酸「ACES」;
1,1−シクロヘキサン二酢酸;
3,6−エンドメチレン−1,2,3,6−テトラヒドロフタル酸「EMTA」(「ENDCA」);
イミダゾール;
塩化2−(アミノエチル)トリメチルアンモニウム「CHOLAMINE」;
N,N−ビス(2−ヒドロキシエチル)−2−アミノエタンスルホン酸「BES」;
2−メチルプロパン−1,2,3−トリスカルボン酸(β−メチルトリカルバリル酸);
2−(N−モルホリノ)プロパン−スルホン酸「MOPS」;
リン酸;
N−トリス(ヒドロキシメチル)メチル−2−アミノエタンスルホン酸「TES」;および
N−2−ヒドロキシエチルピペラジン−N’−2−エタンスルホン酸「HEPES」
から成る群から選択される酸または塩基である、請求項1に記載の組成物。 - 少なくとも1種の生物学的緩衝剤のpKaが5.6〜7.6の範囲である、請求項1に記載の組成物。
- ジチオトレイトールが2mM〜10mMの範囲の濃度で存在する、請求項1に記載の組成物。
- さらに少なくとも1種の生物学的洗剤を、非イオン的相互作用により引き起こされる、抗−HCV抗体以外の抗体のHCV抗原に対する非特異的結合を減少させるのに十分な濃度で含む、請求項1に記載の組成物。
- 少なくとも1種の生物学的洗剤が、0.01%〜1%(%は、g/100mlを意味する)の範囲の濃度で存在する、請求項6に記載の組成物。
- 少なくとも1種の生物学的洗剤が、非イオン界面活性剤、陰イオン界面活性剤、両イオン界面活性剤および陽イオン界面活性剤から成る群から選択される、請求項6に記載の組成物。
- 少なくとも1種の生物学的洗剤が、非イオン界面活性剤である、請求項8に記載の組成物。
- さらに陰イオン界面活性剤を含む、請求項9に記載の組成物。
- さらに、正および負の対イオン源の少なくとも1種を、イオン的相互作用により引き起こされる、抗−HCV抗体以外の抗体のHCV抗原に対する非特異的結合を減少させるのに十分な濃度で含む、請求項1に記載の組成物。
- 少なくとも1種の正および負の対イオン源が、0.05M〜0.5Mの範囲の濃度で存在する、請求項11に記載の組成物。
- 少なくとも1種の正および負の対イオン源が塩である、請求項11に記載の組成物。
- 塩が、NaCl及びKClから成る群から選択される、請求項13に記載の組成物。
- さらに少なくとも1種の粘度調節剤を、微粒子が懸濁した溶液の粘度を増加させて、懸濁した該微粒子の沈降を減少させるのに十分な濃度で含む、請求項1に記載の組成物。
- 少なくとも1種の粘度調節剤が、5%〜25%(%は、g/100mlを意味する)の範囲の濃度で存在する、請求項15に記載の組成物。
- 少なくとも1種の粘度調節剤が糖である、請求項15に記載の組成物。
- 糖が、ショ糖、グルコース及びマンニトールから成る群から選択される、請求項17に記載の組成物。
- さらに少なくとも1種の保存剤を、微生物の増殖を低下させるのに十分な濃度で含む、請求項1に記載の組成物。
- 少なくとも1種の保存剤が、0.1%〜1.0%(%は、g/100mlを意味する)の範囲の濃度で存在する、請求項19に記載の組成物。
- 少なくとも1種の保存剤がアジ化ナトリウムである、請求項19に記載の組成物。
- 組成物のpHが6.4〜6.8の範囲である、請求項1に記載の組成物。
- 熱ストレス条件下における、ウイルスゲノムのNS3領域のC型肝炎ウイルス抗原に係る免疫応答性の安定化に適当な水性組成物であって、
10mM〜500mMの少なくとも1種の生物学的緩衝剤;
2mM〜10mMのジチオトレイトール;
4%〜8%(%はg/100mlを意味する。以下同じ)ののエチレングリコール;
0.01%〜1%の少なくとも1種の生物学的洗剤;
0.05M〜0.5Mの少なくとも1種の正および負の対イオン源;
5%〜25%の少なくとも1種の粘度調整剤;および
0.1%〜1%の少なくとも1種の保存剤を含み、
該組成物のpHは、6.4〜7.2の範囲の濃度である、熱ストレス条件下における、ウイルスゲノムのNS3領域のC型肝炎ウイルス抗原に係る免疫応答性の安定化が可能である、該組成物。
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DK1021719T3 (da) | 1997-09-22 | 2007-06-18 | Novartis Vaccines & Diagnostic | Fremgangsmåde til påvisning af antistoffer i en pröve |
ES2316171T3 (es) | 1997-09-22 | 2009-04-01 | Novartis Vaccines And Diagnostics, Inc. | Tampones para estabilizar antigenos hcv. |
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DE69623303D1 (de) | 2002-10-02 |
JP4101874B2 (ja) | 2008-06-18 |
DE69623303T2 (de) | 2003-04-17 |
JP2001516440A (ja) | 2001-09-25 |
EP0852009A1 (en) | 1998-07-08 |
EP0852009B1 (en) | 2002-08-28 |
WO1996041164A1 (en) | 1996-12-19 |
US5616460A (en) | 1997-04-01 |
US5773212A (en) | 1998-06-30 |
ATE223046T1 (de) | 2002-09-15 |
ES2184866T3 (es) | 2003-04-16 |
JP3930519B2 (ja) | 2007-06-13 |
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