JP2020509076A - 高負荷かつ耐アルカリ性のプロテインa磁性ビーズ及びその使用方法 - Google Patents
高負荷かつ耐アルカリ性のプロテインa磁性ビーズ及びその使用方法 Download PDFInfo
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- JP2020509076A JP2020509076A JP2019556418A JP2019556418A JP2020509076A JP 2020509076 A JP2020509076 A JP 2020509076A JP 2019556418 A JP2019556418 A JP 2019556418A JP 2019556418 A JP2019556418 A JP 2019556418A JP 2020509076 A JP2020509076 A JP 2020509076A
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- alkali
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Abstract
Description
本願において言及される磁性ビーズは、主に四酸化三鉄(Fe3O4)からなる。つまり、四酸化三鉄の量は50%以上である。磁性コアは、機械的研磨法、沈殿法(化学的共沈法、酸化沈殿法、還元沈殿法)、マイクロエマルション法、ソルボサーマル法、ゾルゲル法、有機物熱分解、及び他の方法によって調製することができる。
実施例1で調製した磁性コアは超常磁性である。つまり、外部磁場の作用下で容易に磁化するが、ヒステリシスがなく、ある特定の条件下で化学安定性を有する。しかしながら、この磁性コアは容易に酸化してその超常磁性を失い、その溶液中の分散性は不十分である。酸化を阻止し、強酸及び強塩基に耐える能力を達成するためには、磁性コアを他の材料で被覆する必要がある。この実施例では、実施例1で調製した磁性コアが1つ以上のシリカ層で被覆されることを記載する。
アガロース被覆磁性ビーズは、それぞれ、有機相としてのシクロヘキサン、Span80(SP80)、及び二重蒸留水、乳化剤、ならびに水相を使用した、逆相懸濁法によって調製される。
耐アルカリ性プロテインAなどの他のリガンドが実施例3で調製した磁性ビーズの表面に化学的カップリングによって結合できるようにするには、リガンドへのカップリングを達成するために、例えば、磁性ビーズの表面上のヒドロキシル基のエポキシ活性化により、磁性ビーズの表面を化学的に活性化させることが必要である。100mLのアガロース被覆磁性ビーズを1Lの三角フラスコに加え、1M NaOH溶液を同時に調製した。アガロース被覆磁性ビーズのNaOH溶液に対する体積比が1:1〜1:1.5になるように、1M NaOH溶液を三角フラスコに加え、この混合物を均一に振盪した。最後に、適量のエピクロロヒドリンを1Lの三角フラスコに加え、三角フラスコを振盪インキュベータに入れ、温度を37℃に調整し、反応を1〜1.5時間持続させた。反応の最後に、表面が活性化されたアガロースビーズを得ることができた。
耐アルカリ性プロテインAは、アミノ基、カルボキシル基、ヒドロキシル基、またはスルフヒドリル基を介した共有結合により、活性化されたアガロース磁性ビーズにカップリングされ得る。例えば、耐アルカリ性プロテインAを含むアガロース磁性ビーズは、耐アルカリ性プロテインA中の窒素含有基を有するアミノ酸をエポキシ活性化アガロースの表面に結合させることにより調製され得る。
実施例5で調製した高負荷かつ耐アルカリ性のプロテインAアガロース磁性ビーズを試験した。上記の磁性ビーズを均一に混合し、0.4mLを15mLの遠心管に入れ、磁石(天然の永久磁石または電磁石)を使用し、約30〜60秒にわたって管壁内側に磁性ビーズを吸着させた。上清を捨てたか、またはピペットで除去した。磁石を外し、2mLの二重蒸留水を遠心管に加え、十分に混合してビーズを浄化した。磁性ビーズを磁石によって吸着して管壁内側に定着させた。吸着時間は約30〜60秒であった。上清を捨てたか、またはピペットで除去した。このプロセスを2〜3回繰り返して残りのエタノールを除去した。同様に、磁性ビーズを10mlの20mMリン酸緩衝液で2〜3回浄化し、緩衝液を除去した。
免疫グロブリンIgGに結合した2つのドメインを含むプロテインA、及び免疫グロブリンIgGに結合した5つのドメインを含むプロテインAを、上記のカップリング方法によってそれぞれアガロース磁性ビーズにカップリングした。沈殿した磁性ビーズを各種100μl取り、2mLの二重蒸留水と十分に混合して磁性ビーズを浄化した。5mg/mLのヒトIgG(hIgG)2mLを20mM PBSに溶解させ、前もって浄化した磁性ビーズと共に室温で1時間インキュベートした。次に、磁性ビーズを磁石によって吸着して管壁内側に定着させ、上清を除去した。磁性ビーズを10mLの20mMリン酸緩衝液で2〜3回浄化して、未吸着試料及び不純物を除去した。最後に、目的のタンパク質を500μLの0.1Mグリシン溶出液(pH3.0)で3回溶出した。各溶離液中のhIgGの量を測定して、磁性ビーズの単位体積あたりの静的結合負荷を求めた。結果を以下の表に示す。2つのドメインを含むプロテインA磁性ビーズの負荷は、5つのドメインを含むプロテインA磁性ビーズの負荷よりも高い。
上記の実施例5で調製した耐アルカリ性プロテインAアガロース磁性ビーズを、アルカリ溶液中インサイチュ浄化によって試験した。まず、実施例6の手順に従って免疫グロブリンの精製を行った。pH3.0の0.1Mグリシン溶離液を用いて免疫グロブリンを溶出した後、インサイチュ浄化用アルカリ溶液としての0.5M NaOH溶液5mLに磁性ビーズを15分間浸漬してから、10mLの20mMリン酸緩衝液(0.15M NaCl、30mM Na2HPO4、10mM NaH2PO4を含む。pH7.0)を使用して3回浄化及び平衡化し、1試験サイクルのアルカリ溶液中インサイチュ浄化を完了した。耐アルカリ性プロテインA磁性ビーズの免疫グロブリン結合能力は、各サイクルにおいて、溶離液中の免疫グロブリンの量に基づいて決定することができる。
Claims (19)
- pH2〜14で化学安定性を維持することができ、50mg/mL超の免疫グロブリンIgG結合能を有する、高負荷かつ耐アルカリ性のプロテインA磁性ビーズ。
- 前記磁性ビーズが、pH10〜14のアルカリ溶液中で各回15分間にわたり50回超インサイチュ浄化した後に40mg/mL超の免疫グロブリンIgG結合能を有する、請求項1に記載の高負荷かつ耐アルカリ性のプロテインA磁性ビーズ。
- 前記磁性ビーズが、60eum/g超の比飽和磁化を有する、請求項1に記載の高負荷かつ耐アルカリ性のプロテインA磁性ビーズ。
- 前記磁性ビーズが、20nm〜200nmまたは30μm〜200μmの範囲の粒径を有する、請求項1〜3のいずれか1項に記載の高負荷かつ耐アルカリ性のプロテインA磁性ビーズ。
- 前記磁性ビーズが磁性コア部分及びリガンド部分を含み、前記磁性コア部分の主成分がFe3O4であり、前記リガンド部分が耐アルカリ性プロテインAである、請求項1に記載の高負荷かつ耐アルカリ性のプロテインA磁性ビーズ。
- 前記磁性コア部分がFe2O3を更に含み、Fe2O3:Fe3O4の質量比が1:1〜1:100である、請求項5に記載の高負荷かつ耐アルカリ性のプロテインA磁性ビーズ。
- 前記高負荷かつ耐アルカリ性のプロテインA磁性ビーズ上の前記耐アルカリ性プロテインAの量が3mg/mL以上である、請求項5に記載の高負荷かつ耐アルカリ性のプロテインA磁性ビーズ。
- 前記磁性コア部分が超常磁性である、請求項5または6に記載の高負荷かつ耐アルカリ性のプロテインA磁性ビーズ。
- 前記磁性コア部分が、シリカ、グルカン、アガロース、ポリスチレン、ポリグリシジルメタクリレート、ポリヒドロキシエチルメタクリレート、ポリスチレン−グリシジルメタクリレート、及びそれらの組み合わせのうちの1つ以上から選択される無機材料または有機材料からなる被覆層で被覆されており、前記リガンド部分が前記被覆層にカップリングしている、請求項5に記載の高負荷かつ耐アルカリ性のプロテインA磁性ビーズ。
- 前記被覆層が、前記リガンドと架橋するために必要とされる反応基、または前記被覆層の表面上の化学活性化もしくはカップリングによって前記リガンドと架橋するために必要とされる反応基を有する、請求項9に記載の高負荷かつ耐アルカリ性のプロテインA磁性ビーズ。
- 前記反応基が、ヒドロキシル基、カルボキシル基、アミノ基、及びエポキシ基から選択される、請求項10に記載の高負荷かつ耐アルカリ性のプロテインA磁性ビーズ。
- 前記耐アルカリ性プロテインAが、過酷なアルカリ性条件下で処置された後に免疫グロブリンIgGに確実に結合できるように、pH10〜14の強アルカリ環境においてタンパク質高次構造の安定性を維持することができる、請求項5に記載の高負荷かつ耐アルカリ性のプロテインA磁性ビーズ。
- 前記耐アルカリ性プロテインAが、免疫グロブリンIgGに結合することができる2〜4つのドメインを含む、請求項5に記載の高負荷かつ耐アルカリ性のプロテインA磁性ビーズ。
- 前記耐アルカリ性プロテインAが、前記アガロースにカップリングすることにより前記被覆層に結合している、請求項5に記載の高負荷かつ耐アルカリ性のプロテインA磁性ビーズ。
- 前記耐アルカリ性プロテインAが、配列番号1のアミノ酸配列または配列番号2のアミノ酸配列を含む、請求項5に記載の高負荷かつ耐アルカリ性のプロテインA磁性ビーズ。
- 前記耐アルカリ性プロテインAが、前記アミノ酸配列の相同2〜4量体及び/または異種2〜4量体である、請求項15に記載の高負荷かつ耐アルカリ性のプロテインA磁性ビーズ。
- 前記耐アルカリ性プロテインAが、組換え発現された融合タンパク質の形態である、請求項16に記載の高負荷かつ耐アルカリ性のプロテインA磁性ビーズ。
- 免疫グロブリンを精製及び/または検出するための方法であって、請求項1〜17のいずれか1項に記載の高負荷かつ耐アルカリ性のプロテインA磁性ビーズに、前記免疫グロブリンを含む試料を接触させるステップを含む、前記方法。
- 請求項1〜17のいずれか1項に記載の高負荷かつ耐アルカリ性のプロテインA磁性ビーズを再生するための方法であって、前記高負荷かつ耐アルカリ性のプロテインA磁性ビーズを、0.1〜1時間にわたり、0.1M〜0.5Mの水酸化ナトリウム溶液もしくは水酸化カリウム溶液またはこれら両方の混合溶液に浸漬してから、前記磁性ビーズを純水もしくは緩衝液に浸すか、または前記磁性ビーズを3〜5回すすいで、前記アルカリ溶液を完全に除去することと、前記高負荷かつ耐アルカリ性のプロテインA磁性ビーズを平衡化緩衝液中で保存することとを含む、前記方法。
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Citations (4)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
JP2014207905A (ja) * | 2009-03-10 | 2014-11-06 | 独立行政法人産業技術総合研究所 | 膜に提示したタンパク質を特異的に認識するポリペプチドの調製方法 |
CN104475041A (zh) * | 2014-10-22 | 2015-04-01 | 哈尔滨工业大学 | 制备琼脂糖磁性微球的新方法及其在分离纯化IgG抗体中的用途 |
WO2016104338A1 (ja) * | 2014-12-22 | 2016-06-30 | 和光純薬工業株式会社 | ビオチン化核酸の分離方法 |
US20160237124A1 (en) * | 2013-03-18 | 2016-08-18 | Hong Qian | Kind of mutated proteins a with high alkali resistance feature and application thereof |
Family Cites Families (20)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
US4452773A (en) * | 1982-04-05 | 1984-06-05 | Canadian Patents And Development Limited | Magnetic iron-dextran microspheres |
AU7685498A (en) * | 1997-05-14 | 1998-12-08 | Weixin Tang | Fully-coated, uniform-sized metallic particles |
WO2005076938A2 (en) | 2004-02-11 | 2005-08-25 | Massachusetts Institute Of Technology | Multi-polymer-coated magnetic nanoclusters |
JP2005312353A (ja) * | 2004-04-28 | 2005-11-10 | Hitachi Maxell Ltd | 磁気ビーズ及びこれを用いた検体の分離方法 |
US7897257B2 (en) | 2005-04-18 | 2011-03-01 | Ge Healthcare Bio-Sciences Ab | Magnetic beads comprising an outer coating of hydrophilic porous polymer and method of making thereof |
WO2007050017A1 (en) | 2005-10-28 | 2007-05-03 | Ge Healthcare Bio-Sciences Ab | Separation medium with various functionalities |
CN1975423A (zh) * | 2006-09-06 | 2007-06-06 | 浙江清华长三角研究院 | 免疫磁珠及制作方法和用于检测的方法及测试板 |
KR101092916B1 (ko) * | 2007-09-05 | 2011-12-12 | (주) 에이프로젠 | 중간엽줄기세포 분리 동정 바이오 마커 발굴 및 이를이용한 중간엽줄기세포 분리 방법 |
CN100593545C (zh) * | 2007-10-23 | 2010-03-10 | 北京博迈世纪生物技术有限公司 | 一种免疫纳米磁性葡聚糖微球的制备方法 |
EP2412809B1 (en) * | 2009-03-24 | 2017-08-09 | Kaneka Corporation | Protein having affinity for immunoglobulin, and immunoglobulin-binding affinity ligand |
CN102838674A (zh) * | 2012-09-20 | 2012-12-26 | 海狸纳米科技(苏州)有限公司 | 一种利用蛋白a的高密度包被磁珠纯化抗体的方法 |
WO2014064238A1 (en) * | 2012-10-25 | 2014-05-01 | Affibody Ab | Method for the separation of proteins containing an albumin-binding |
CN103007846B (zh) * | 2012-12-14 | 2015-02-04 | 无锡百运纳米科技有限公司 | 一种载蛋白的磁性微球的制备方法 |
GB201301631D0 (en) * | 2013-01-30 | 2013-03-13 | Ge Healthcare Ltd | Separation of substances using magnetic beads |
CN103212377B (zh) * | 2013-04-19 | 2014-09-24 | 哈尔滨益材新材料有限公司 | 一种琼脂糖免疫磁性微球的制备方法及其应用 |
CN103275217A (zh) * | 2013-06-26 | 2013-09-04 | 河南工业大学 | 磁珠法快速纯化抗体 |
CN103483626B (zh) * | 2013-09-16 | 2015-12-02 | 海狸纳米科技(苏州)有限公司 | 一种磁性琼脂糖微球 |
CN103483602B (zh) | 2013-09-16 | 2015-07-22 | 海狸纳米科技(苏州)有限公司 | 一种小粒径磁性琼脂糖微球的制备方法 |
CN105600832A (zh) * | 2014-11-19 | 2016-05-25 | 正大天晴药业集团股份有限公司 | 一种经修饰的超顺磁性氧化铁的制备方法 |
CN105418730A (zh) * | 2015-11-13 | 2016-03-23 | 洛阳惠尔纳米科技有限公司 | 一种基于磁珠法的抗体纯化方法 |
-
2018
- 2018-01-04 WO PCT/CN2018/071397 patent/WO2018127099A1/zh unknown
- 2018-01-04 JP JP2019556418A patent/JP2020509076A/ja active Pending
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Patent Citations (4)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
JP2014207905A (ja) * | 2009-03-10 | 2014-11-06 | 独立行政法人産業技術総合研究所 | 膜に提示したタンパク質を特異的に認識するポリペプチドの調製方法 |
US20160237124A1 (en) * | 2013-03-18 | 2016-08-18 | Hong Qian | Kind of mutated proteins a with high alkali resistance feature and application thereof |
CN104475041A (zh) * | 2014-10-22 | 2015-04-01 | 哈尔滨工业大学 | 制备琼脂糖磁性微球的新方法及其在分离纯化IgG抗体中的用途 |
WO2016104338A1 (ja) * | 2014-12-22 | 2016-06-30 | 和光純薬工業株式会社 | ビオチン化核酸の分離方法 |
Non-Patent Citations (1)
Title |
---|
JOURNAL OF MAGNETISM AND MAGNETIC MATERIALS, vol. 293, JPN6021042544, 2005, pages 345 - 348, ISSN: 0004812283 * |
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EP3578255A4 (en) | 2021-01-13 |
US20190329218A1 (en) | 2019-10-31 |
WO2018127099A1 (zh) | 2018-07-12 |
US11577218B2 (en) | 2023-02-14 |
EP3578255B1 (en) | 2022-06-22 |
CA3048273A1 (en) | 2018-07-12 |
CN110177617A (zh) | 2019-08-27 |
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