JP2022520024A - トラックエッチングされた膜を用いた生物学的流体の濾過方法 - Google Patents
トラックエッチングされた膜を用いた生物学的流体の濾過方法 Download PDFInfo
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- JP2022520024A JP2022520024A JP2021543358A JP2021543358A JP2022520024A JP 2022520024 A JP2022520024 A JP 2022520024A JP 2021543358 A JP2021543358 A JP 2021543358A JP 2021543358 A JP2021543358 A JP 2021543358A JP 2022520024 A JP2022520024 A JP 2022520024A
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- filtrate
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Abstract
Description
TE膜は、細胞培養物の濾過およびその他の生物学的分離に適していることがここで判明した。いかなる理論にも縛られることを望むものではないが、これらの膜の性能は、汚れに対する耐性、および微細孔の形状とサイズを制御する能力によるものであり、その結果、効率的かつ効果的な分離が可能になると考えられている。これらの特性は、多くのバイオプロセス応用に有利であると考えられる。
本開示のある原則は、以下の実施例によってさらに説明される。
フラットシート状のTE膜を、「Eスクリーン」スペーサ(すなわち、ウォルサムMA、Repligen Corporation社製の粗い供給スペーサ)を用いて100cm2のカセットに作製した。カセットを純水で洗浄し、流路を空にして、カセットが一体化して使用に適しているかどうかを試験するために、一体性空気試験を実行した。MF膜の典型的な試験条件範囲である3psiでは、空気流は1.2ml/min未満であるため、カセットが一体化していることが分かった。
微細孔径0.2ミクロンのフラットシート状のTE膜を、「Eスクリーン」(すなわち、Repligen社製の粗い供給スペーサ)を用いて100cm2のカセットに作製した(TE02)。その性能を、2種類のチャイニーズハムスター卵巣(CHO)細胞培養供給液の流れを用いて、0.2ミクロンのポリエーテルスルホン(PES)位相反転膜をスキンアップ(PES02SU)およびスキンダウン(PES02SD)方向に使用したカセットと比較した。その結果、図2のような良好な結果が得られた。TE膜は、同程度の微細孔径のPES膜と比較して、より高い透過性(LMH/psi)、より高い容量(L/m2)およびより高い篩い分け性を示した。TE02膜のタンパク質の篩い分け率は有意に高く、78%で一定していた。PES02SUでは、開始時に68%だった篩い分け率が終了時には34%と、すなわちTE02カセットで得られた篩い分け率の半分以下にまで急激に低下した。
TFF応用に必要な圧力にTE膜で作られたカセットが耐えられるかどうかを試験するために、100cm2のTEカセットに水を通し、様々な膜間圧力(TMP)で水流束を測定した。試験は、TMP値を1~8.5psiのようにすなわち低い値から高い値へと段階的に増加させ、各TMPでの水流束を測定して行われた。高圧で試験しても膜が損傷しないように、TMPを8.5psiから2.2psiおよび1.2psiと高値から低値に下げて試験を実施した。図3に示すように、どちらの方向に測定した水流束にも大きな差異はなく、圧力の安定性が強固であることが示されている。
TE膜を使用したカセットの化学的安定性を試験するため、100cm2のカセットを0.2Nの水酸化ナトリウム溶液で、膜面積1m2あたり2リットル使用して洗浄した。カセットを30分間洗浄し、3psiのTMPで前後の水流束を測定した。図4に示すように、化学洗浄前後の水流束はほぼ同じであり、カセットの洗浄や保管に一般的に使用されている試薬に対する化学的安定性が高く、再使用可能なカセットの製造が可能であることを示している。
Claims (23)
- 可撓性隔離板と、
ガスケットおよび濾板の少なくとも1つと、
前記可撓性隔離板と前記ガスケットまたは前記濾板との間に配置され、少なくとも1つの膜、少なくとも1つの供給流路、および少なくとも1つの濾液流路を画定する、複数のインターリーブ層と
を備える接線流濾過カセットであって、
前記複数のインターリーブ層は、
1または複数の流体ポートを含み、内周で囲まれた開放的な内部容積を画定する濾液流路スペーサと、
内周で囲まれた開放的な内部容積を画定し、1または複数の流体ポートを含む非可撓性の供給流路スペーサと、
前記濾液流路スペーサと前記供給流路スペーサとの間に配置された膜と、任意に、
前記膜と、前記濾液流路スペーサおよび前記供給流路スペーサの一方とを接着する感圧性接着剤であって、前記感圧性接着剤の薄膜は、隣接する流路の高さの50%未満の厚さを有する、感圧性接着剤と
を含み、
(a)前記可撓性隔離板は、可撓性ポリマーまたは熱可塑性エラストマーを含み、インターリーブスタックの第1表面に接着されており、(b)前記膜は、複数のトラックエッチングされた膜(TE膜)を含み、前記TE膜は複数の微細孔を含み、前記微細孔の孔径および前記微細孔の形状が均一である
接線流濾過(TFF)カセット。 - 前記複数の微細孔は、直径、深さ、および/または経路長が均一である、請求項1に記載のTFFカセット。
- 平均微細孔径が約100kD~約30ミクロンである、請求項1または2に記載のTFFカセット。
- 前記TE膜が60ミクロンまでの厚さを有する、請求項1から3のいずれか一項に記載のTFFカセット。
- 前記トラックエッチングされた膜が、複数のポリカーボネート、ポリイミド、ポリフッ化ビニリデン、またはポリエステルのフラットシートからなる、請求項1から4のいずれか一項に記載のTFFカセット。
- 前記複数の微細孔は、円筒形、円錐形、葉巻形、漏斗形および砂時計形のうちの1または複数の構造を有してよい、請求項1から5のいずれか一項に記載のTFFカセット。
- 前記トラックエッチングされた膜が滑らかな表面を有する、請求項1から6のいずれか一項に記載のTFFカセット。
- 前記接線流濾過カセットが入れ替わる接線流濾過に使用される、請求項1から7のいずれか一項に記載のTFFカセット。
- 前記インターリーブスタックの第2表面にガスケットが接着されており、前記ガスケットは、供給流路と濾液流路のための別々の流体ポートを備える、請求項1から8のいずれか一項に記載のTFFカセット。
- 前記ガスケットには濾板が接着されており、前記濾板は、前記濾板の供給ポートが前記ガスケットの供給ポートと整列し、前記濾板の濾液ポートが前記ガスケットの濾液ポートと整列する流体マニホールドを備える、請求項1から9のいずれか一項に記載のTFFカセット。
- 在庫管理単位(SKU)番号、ロット番号、シリアル番号、容量、供給および/または濾液流路の数、および膜の数のうちの1または複数によって前記TFFカセットを識別するために、前記接線流濾過カセットの側壁にタブまたは刻印をさらに備える、請求項1から10のいずれか一項に記載のTFFカセット。
- 前記タブまたは刻印はバーコードからなる、請求項11に記載のTFFカセット。
- 前記接線流濾過カセットが使用後に使い捨て可能である、請求項1から12のいずれか一項に記載のTFFカセット。
- 前記TFFカセットが再使用可能である、請求項1から13のいずれか一項に記載のTFFカセット。
- 前記TFFカセットが密封されたエッジを備える、請求項1から14のいずれか一項に記載のTFFカセット。
- 前記少なくとも1つの供給流路は、前記供給流路スペーサによって画定される空間内に配置された供給スクリーンを備える、請求項1から15のいずれか一項に記載のTFFカセット。
- 前記供給スクリーンが、織布、不織布または押出ポリマーメッシュからなる、請求項16に記載のTFFカセット。
- 前記少なくとも1つの濾液流路は、前記濾液流路スペーサによって画定された空間内に配置された濾液スクリーンを備える、請求項16に記載のTFFカセット。
- 前記濾液スクリーンが、織布、不織布、または押出ポリマーメッシュからなる、請求項18に記載のTFFカセット。
- カセットを使用する段階であって、
前記カセットは、
可撓性隔離板と、
ガスケットおよび濾板の少なくとも1つと、
前記可撓性隔離板と前記ガスケットまたは前記濾板との間に配置され、少なくとも1つの供給流路および少なくとも1つの濾液流路を画定する、複数のインターリーブ層と
を含み、
前記複数のインターリーブ層は、
1または複数の流体ポートを含み、内周で囲まれた開放的な内部容積を画定する濾液流路スペーサと、
内周で囲まれた開放的な内部容積を画定し、1または複数の流体ポートを含む非可撓性の供給流路スペーサと、
前記濾液流路スペーサと前記供給流路スペーサとの間に配置された膜と、任意に、
前記膜と、前記濾液流路スペーサおよび前記供給流路スペーサの一方とを接着する感圧性接着剤であって、前記感圧性接着剤の薄膜は、隣接する流路の高さの50%未満の厚さを有する、感圧性接着剤と
を含み、
(a)前記可撓性隔離板は、可撓性ポリマーまたは熱可塑性エラストマーを含み、インターリーブスタックの第1表面に接着されており、(b)前記膜は、複数のトラックエッチングされた膜を含み、前記トラックエッチングされた膜は複数の微細孔を含み、前記微細孔の孔径および前記微細孔の形状が均一である、
接線流濾過カセットである
段階と、
カセットハウジングをプロセス容器に接続する段階と、
前記カセットハウジングを分離システムに接続する段階と、
前記トラックエッチングされた膜フィルタと前記分離システムに細胞培養物を循環させる段階と、
前記細胞培養物を前記膜フィルタで濾過する段階と、
得られた濾液を収集する段階と、
前記細胞培養物を前記プロセス容器に戻す段階と
を備える、細胞培養物を濾過する方法。 - 前記細胞培養物が哺乳類の細胞培養物である、請求項20に記載の方法。
- 前記細胞培養物の循環が、入れ替わる接線流によって実行される、請求項20に記載の方法。
- 前記細胞培養物の循環が、接線流濾過によって実行される、請求項20に記載の方法。
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