JP7044222B2 - 実質的にコアギュローゲンを含まないリムルスアメボサイトライセートを使用してエンドトキシンを検出する方法 - Google Patents
実質的にコアギュローゲンを含まないリムルスアメボサイトライセートを使用してエンドトキシンを検出する方法 Download PDFInfo
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Description
基質+H2O+酵素→ペプチド+pNA(黄色)
[0049]いくつかの実施形態において、グラム陰性菌エンドトキシンは、LAL中の酵素前駆体の活性化を間接的に触媒することが可能である。活性化の最初の速度は、存在するエンドトキシンの濃度によって決まる可能性がある。
タンジェンシャルフローろ過を使用したLALの精製
[0070]カブトガニの血液細胞(アメボサイト)から抽出したリムルスアメボサイトライセート(LAL)をロンザ(バーゼル、スイス)から入手した。コアギュローゲンを除去するために、タンジェンシャルフローろ過(TFF)用のホローファイバー修飾ポリエーテルスルホン(mPES)メンブランフィルター(スペクトラムラボラトリーズ(SpectrumLabs))にLALを供した。30kDa MWCO mPESフィルター、コールパーマー(Cole Parmer)マスターフレックスポンプチュービング(Masterflex Pump Tubing)並びに適したポンプ及び背圧弁を使用してTFFシステムをセットアップした。そのシステムをLAL試薬水(LRW)で洗い流し、その後、1N NaOHを使用して室温で1時間脱パイロジェン処理をした。脱パイロジェン処理後、LRWを使用して、システムをすすぎ、その後、そのシステムを、TFFバッファーを使用して平衡化した。
コアギュローゲンの定量
[0074]LAL保持液に対するSDS-PAGE/タンパク質ゲル染色を実施し、元のLAL中の20kDコアギュローゲンバンドのバンド密度を、あれば、TFF保持液中のコアギュローゲンバンドのバンド密度と比較することによって、LAL中の残存コアギュローゲンの半定量的評価を行った。例えば、図7A参照。
実質的にコアギュローゲンを含まないLALの安定性
[0079]実質的にコアギュローゲンを含まないLALの安定性を調査した。実質的にコアギュローゲンを含まないLALサンプルを1、2、3、4、5、6、7、8、9及び10週間保管した。示した期間の終わりに、実質的にコアギュローゲンを含まないLALサンプルを96ウェルプレート中のLAL発色試薬と混合し、405nmにおける吸光度を測定するインキュベートプレートリーダーに入れる。その反応を黄色の外観により経時的に自動的に監視した。
コアギュローゲン除去は分離を改善する
[0081]複数のオンセットmODにおける分離に対して、LALからコアギュローゲンを除去することが有する効果の調査を行った。通常のLAL対実質的にコアギュローゲンを含まないLALの比較は、実質的にコアギュローゲンを含まないLALを使用した場合に、それぞれのオンセットmODにおける分離の増加を示した。分離を増加させるために実質的にコアギュローゲンを含まないLALを使用することによって、アッセイの目標の感度を、より低いオンセットmOD設定においてより短い時間で達成することができる。配合物を調製するために実質的にコアギュローゲンを含まないLALを使用すると、0.005EU/mlの目標の感度を、368秒の分離で26分で達成できる。通常のLALを使用すると、0.005EU/mlの感度を50mODにおいて32分で達成するが、分離はわずか67秒である。
LAL及び実質的にコアギュローゲンを含まないLAL配合物の発色検出
[0083]LAL及び実質的にコアギュローゲンを含まないLALの両方のサンプルを、0.005EU/ml、0.05EU/ml、0.5EU/ml、5EU/ml及び50EU/mlの標準に対してLAL調製物中20%、30%、40%及び50%の濃度で試験した。さらに、さまざまな濃度のズイッタージェントも試験した。図3及び図4。このデータは、実質的にコアギュローゲンを含まないLAL及びズイッタージェントの濃度を高めると、LAL及び実質的にコアギュローゲンを含まないLAL配合物の両方に関してそれぞれ標準に対する反応速度が増加したことを示唆している。但し、LAL配合物により反応速度が増加するため、ブランクと最低の標準(0.005EU/ml)との間では分離が失われる一方で、すべての実質的にコアギュローゲンを含まないLAL配合物に関しては、少なくとも200秒の分離が維持されている。増加したズイッタージェントと配合した実質的にコアギュローゲンを含まないLALは、許容される分離で0.001EU/mlの感度を達成した。(データは示していない)。
実質的にコアギュローゲンを含まないLALと発色基質の比率
[0084]実質的にコアギュローゲンを含まないLALと発色基質の最適な比率を200mOD及び50mODにおいて調査した。図5A及び図5B。このデータは、70%のLAL調製物と30%の発色基質の比率が最速の反応時間になったことを示している。実質的にコアギュローゲンを含まないLALの3つの異なるロットを見ると、この観察結果が確認された。図6A。
本出願は、ASCII形式で電子的に提出された配列表を含有し、参照によりその全体が本明細書に組み込まれる。2017年9月5日に作成された上記ASCIIコピーは、0132-0023WO1_SL.txtという名前であり、1,055バイトのサイズである。
Claims (20)
- 発色アッセイを使用して、サンプル中のエンドトキシンを検出する方法であって、前記方法が、
a.前記サンプルをリムルスアメボサイトライセート(LAL)及び発色基質を含む試薬と接触させるステップと、
b.前記サンプル中のエンドトキシンの存在下における前記発色基質の変化によって生じる発色効果を測定するステップとを含み、
前記LALが、タンパク質染色を伴うSDS-PAGEにより測定される場合に、LAL中の総タンパク質に対して5%(wt/wt)未満のコアギュローゲンを含むものであり、
前記方法は、0.001EU/mL又は0.001EU/mL未満の感度を有する、方法。 - 前記発色基質の変化が、酵素反応により起こり、前記酵素反応が、ポリペプチドからの発色団の切断である、請求項1に記載の方法。
- 前記試薬が、水性液体である、請求項1又は2に記載の方法。
- 前記LAL、発色基質又はその両方が、凍結乾燥され、その後、前記サンプルと接触させるステップに先立って再構成される、請求項1~3のいずれか一項に記載の方法。
- 前記サンプルが、非経口剤形、ワクチン、抗生剤、治療用タンパク質、治療用核酸、治療用抗体、及びバイオテクノロジー製品からなる群から選択される、請求項1~4のいずれか一項に記載の方法。
- 前記LALが、SDS-PAGE及びタンパク質染色によって測定される場合に、前記LAL中の総タンパク質に対して2%(wt/wt)未満のコアギュローゲンを含む、請求項1~5のいずれか一項に記載の方法。
- 前記LALが、タンパク質染色を伴うSDS-PAGEによって測定される場合に、前記LAL中の総タンパク質に対して0.5%(wt/wt)未満のコアギュローゲンを含む、請求項1~6のいずれか一項に記載の方法。
- 発色アッセイを使用して、生物学的サンプル中のエンドトキシンを検出する方法であって、前記方法が、
a.前記生物学的サンプルをリムルスアメボサイトライセート(LAL)及びAc-Ile-Glu-Ala-Arg-pNA(配列番号:1)を含む水性試薬と接触させるステップと、
b.前記サンプル中のエンドトキシンの存在下におけるAc-Ile-Glu-Ala-Arg-pNA(配列番号:1)からのpNAの酵素切断によって生じる405nmにおける吸光度の変化を測定するステップとを含み、
前記LALが、タンパク質染色を伴うSDS-PAGEにより測定される場合に、LAL中の総タンパク質に対して5%(wt/wt)未満のコアギュローゲンを含むものであり、
前記方法は、0.001EU/mL又は0.001EU/mL未満の感度を有する、方法。 - 前記発色基質が、Ac-Ile-Glu-Ala-Arg-pNA(配列番号:1)である、請求項1~7のいずれか一項に記載の方法。
- タンパク質染色を伴うSDS-PAGEにより測定される場合に、LAL中の総タンパク質に対して5%(wt/wt)未満のコアギュローゲンを含むリムルスアメボサイトライセート(LAL)を生成する方法であって、前記方法が、
コアギュローゲンを含むLAL及びバッファーを含む組成物を20kDa~50kDaフィルターを使用したタンジェンシャルフローろ過(TFF)に供するステップであって、前記TFFが保持液及びろ液を生成するステップと、
前記保持液を回収し、それによりLAL中の総タンパク質に対して5%(wt/wt)未満のコアギュローゲンを含むLALを得るステップと
を含む、方法。 - 前記TFFが、350ml/分~600ml/分の流量で実施される、請求項10に記載の方法。
- 前記TFFが、350ml/分~500ml/分の流量で実施される、請求項11に記載の方法。
- LALを含む前記組成物が、バッファーである、請求項10~12のいずれか一項に記載の方法。
- 前記バッファーがトリスバッファー又はMESバッファーである、請求項13に記載の方法。
- 前記バッファーが6.0~9.0のpHを有する、請求項13又は14に記載の方法。
- 前記バッファーが7.0~8.0のpHを有する、請求項15に記載の方法。
- 前記TFFフィルターが、修飾ポリエーテルスルホン(mPES)、ポリスルホン(PS)及びポリエーテルスルホン(PES)から選択される膜を含む、請求項10~16のいずれか一項に記載の方法。
- 前記TFFフィルターが、修飾ポリエーテルスルホン(mPES)メンブランフィルターである、請求項16又は17に記載の方法。
- 前記LALが、タンパク質染色を伴うSDS-PAGEによって測定される場合に、前記LAL中の総タンパク質に対して2%(wt/wt)未満のコアギュローゲンを含む、請求項10~18のいずれか一項に記載の方法。
- 前記LALが、タンパク質染色を伴うSDS-PAGEによって測定される場合に、前記LAL中の総タンパク質に対して0.5%(wt/wt)未満のコアギュローゲンを含む、請求項10~19のいずれか一項に記載の方法。
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Publication number | Priority date | Publication date | Assignee | Title |
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US9458450B2 (en) | 2012-03-15 | 2016-10-04 | Flodesign Sonics, Inc. | Acoustophoretic separation technology using multi-dimensional standing waves |
US10967298B2 (en) | 2012-03-15 | 2021-04-06 | Flodesign Sonics, Inc. | Driver and control for variable impedence load |
US10704021B2 (en) | 2012-03-15 | 2020-07-07 | Flodesign Sonics, Inc. | Acoustic perfusion devices |
US9950282B2 (en) | 2012-03-15 | 2018-04-24 | Flodesign Sonics, Inc. | Electronic configuration and control for acoustic standing wave generation |
WO2015105955A1 (en) | 2014-01-08 | 2015-07-16 | Flodesign Sonics, Inc. | Acoustophoresis device with dual acoustophoretic chamber |
US11021699B2 (en) | 2015-04-29 | 2021-06-01 | FioDesign Sonics, Inc. | Separation using angled acoustic waves |
US11708572B2 (en) | 2015-04-29 | 2023-07-25 | Flodesign Sonics, Inc. | Acoustic cell separation techniques and processes |
US11377651B2 (en) | 2016-10-19 | 2022-07-05 | Flodesign Sonics, Inc. | Cell therapy processes utilizing acoustophoresis |
US11474085B2 (en) | 2015-07-28 | 2022-10-18 | Flodesign Sonics, Inc. | Expanded bed affinity selection |
US11459540B2 (en) | 2015-07-28 | 2022-10-04 | Flodesign Sonics, Inc. | Expanded bed affinity selection |
US11085035B2 (en) | 2016-05-03 | 2021-08-10 | Flodesign Sonics, Inc. | Therapeutic cell washing, concentration, and separation utilizing acoustophoresis |
US11214789B2 (en) | 2016-05-03 | 2022-01-04 | Flodesign Sonics, Inc. | Concentration and washing of particles with acoustics |
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---|---|---|---|---|
JP2014014375A (ja) | 2007-11-12 | 2014-01-30 | Hiroshima Univ | エンドトキシンの濃度測定方法および濃度測定用キット |
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---|---|---|---|---|
US4322217A (en) | 1980-06-27 | 1982-03-30 | Mallinckrodt, Inc. | Process for preparing Limulus lysate |
DK0426395T3 (ja) * | 1989-10-30 | 1997-03-03 | Biowhittaker Inc | |
JP3122454B2 (ja) | 1990-09-27 | 2001-01-09 | 生化学工業株式会社 | カブトガニ・アメボサイト・ライセートの調製法 |
US5208148A (en) | 1990-12-07 | 1993-05-04 | Molecular Probes, Inc. | Lipophilic fluorescent glycosidase substrates |
US5576424A (en) | 1991-08-23 | 1996-11-19 | Molecular Probes, Inc. | Haloalkyl derivatives of reporter molecules used to analyze metabolic activity in cells |
DE69229174T2 (de) | 1991-08-23 | 1999-09-16 | Molecular Probes Inc | Verwendung von haloalkylderivaten von reportermolekülen zur analyse der metabolischen aktivität in zellen |
US5242805A (en) | 1991-08-23 | 1993-09-07 | Molecular Probes, Inc. | Long wavelength lipophilic fluorogenic glycosidase substrates |
JP3242733B2 (ja) | 1993-02-26 | 2001-12-25 | 生化学工業株式会社 | エンドトキシン特異的測定剤 |
US5830912A (en) | 1996-11-15 | 1998-11-03 | Molecular Probes, Inc. | Derivatives of 6,8-difluoro-7-hydroxycoumarin |
US5773236A (en) | 1997-04-25 | 1998-06-30 | Molecule Probes, Inc. | Assay for glutathiane transferase using polyhaloaryl-substituted reporter molecules |
WO2008114149A2 (en) | 2007-03-21 | 2008-09-25 | Id Biomedical Corporation Of Quebec | Chimeric antigens |
US7846678B2 (en) | 2008-08-18 | 2010-12-07 | BioDtech, Inc. | Enhancing endotoxin detection |
MY155541A (en) | 2010-10-29 | 2015-10-30 | Universiti Malaysia Terengganu | Process for the preparation of amoebocyte lysate from the haemolymph of the horseshoe crab |
CN102305788A (zh) | 2010-11-22 | 2012-01-04 | 天津市一瑞生物工程有限公司 | 革兰阴性菌脂多糖(内毒素)比色检测试剂盒的制备及其使用方法 |
SG11201900767VA (en) | 2016-08-03 | 2019-02-27 | Lonza Walkersville Inc | Method of detecting an endotoxin using limulus amebocyte lysate substantially free of coagulogen |
CN110192110B (zh) * | 2017-01-11 | 2023-01-10 | 隆萨沃克斯维尔股份有限公司 | 无凝固蛋白原的澄清的鲎阿米巴样细胞裂解物 |
US20190001330A1 (en) * | 2017-06-28 | 2019-01-03 | Lonza Walkersville, Inc. | Cartridge for endotoxin detection |
-
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Patent Citations (1)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
JP2014014375A (ja) | 2007-11-12 | 2014-01-30 | Hiroshima Univ | エンドトキシンの濃度測定方法および濃度測定用キット |
Non-Patent Citations (3)
Title |
---|
James C. Hurley,Endotoxemia: Methods of detection and clinical correlates,Clinical Microbiology Reviews,1995年01月01日,vol 8, no.2,268-292 |
Shin Nakamura et al.,Fractionation of limulus amebocyte lysate,Biochimica et Biophysica Acta,1982年10月05日,707,217-225 |
Toshie Harada-Suzuki et al.,Further studies on the chromogenic substrate assay method for bacterial endotoxins using horseshoe crab (Tachypleus tridentatus) hemocyte lysate,Journal of Biochemistry,1982年12月01日,92,793-800 |
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SG11201900767VA (en) | 2019-02-27 |
EP3943608B1 (en) | 2023-07-12 |
EP3943608C0 (en) | 2023-07-12 |
KR20190037284A (ko) | 2019-04-05 |
EP3494226A4 (en) | 2020-03-18 |
IL264464B1 (en) | 2023-10-01 |
EP3943608A1 (en) | 2022-01-26 |
WO2018026941A1 (en) | 2018-02-08 |
IL264464B2 (en) | 2024-02-01 |
US20220074940A1 (en) | 2022-03-10 |
IL264464A (en) | 2019-02-28 |
CN109790561B (zh) | 2024-02-06 |
EP3494226A1 (en) | 2019-06-12 |
EP3494226B1 (en) | 2021-09-22 |
CN109790561A (zh) | 2019-05-21 |
US11156614B2 (en) | 2021-10-26 |
US20180038864A1 (en) | 2018-02-08 |
JP2019526052A (ja) | 2019-09-12 |
US11906521B2 (en) | 2024-02-20 |
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