JP7391840B2 - 乳酸塩の蓄積を予防しつつ、細胞培養物を増殖させるための方法及びシステム - Google Patents
乳酸塩の蓄積を予防しつつ、細胞培養物を増殖させるための方法及びシステム Download PDFInfo
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Description
[0001]本出願は、2017年11月20日出願の米国特許仮出願第62/588,464号、及び2018年10月18日出願の米国特許仮出願第62/747,311号に基づき、その優先権を主張するものであり、両号は参考として本明細書に組み込まれている。
[0002]バイオリアクターは、生物学的反応又はプロセスが研究室スケール又は工業スケールで実施可能な装置であり、生物製剤業界において幅広く使用されている。バイオリアクターは、すべての異なる種類のバイオプロダクトを製造するのに使用可能である。バイオプロダクトは、例えば、細胞培養物、及び飲料、バイオ燃料、バイオエネルギー、生化学物質、抗生物質、アミノ酸、酵素、モノクロナール抗体、モノマー、タンパク質、食品培養物、バイオポリマー、アルコール、着香料、芳香剤等を含む、細胞培養物に由来する材料を含み得る。いくつかの実施形態では、細胞培養物は、細胞療法用として増殖され得る。細胞療法は、ex vivoで操作又は改変された自己、同種異系、又は異種の細胞の投与により、ヒトにおける疾患又は傷害を予防、治療、治癒、又は緩和することである。細胞療法の1つの目的は、傷害を受けた組織又は臓器を修復、置き換え、又は復元することである。
[0009]本開示は、バイオマテリアル、例えば細胞培養物等を増殖させるための方法及びシステムを一般的に目的とする。一実施形態では、例えば、本開示の方法及びシステムは、哺乳動物の細胞培養物を増殖させることを目的とする。本開示に従って、強固な品質特性濃度、例えば乳酸塩濃度等を、細胞培養物のインキュベーション期間全体を通じて決定する能力を有する予測モデルを含有するコントローラーが開発された。予測モデルは、品質特性濃度を事前設定された限界内に維持するために、増殖期間中に、細胞培養物内の少なくとも1つの条件を選択的に変更するのに使用可能である。例えば、本開示の方法及びシステムを通じて、細胞培養物は、プロセスの終了時に、細胞培養物内の乳酸塩蓄積を防止する方式で増殖し得る。
[00029]本考察は、例示的な実施形態の説明に過ぎず、また本開示のより広い態様を制限するようには意図されないものと、当業者により理解される。
(式中、y(t)はアウトプット/被コントロール変数であり、uj(t)は被操作変数niのうちの1つを表し、nkは時間遅延であり、naは極の数であり、nbはゼロの数であり、並びにai及びbjiは識別プロセスにより決定される係数である)の形式のものである。時間変動式のARXモデルでは、各パラメーターの影響を表す係数は、モデルが時間変動式であるように、時間(すなわち、日)と共に変化する。(1)に記載されるARXモデルは、ワンステップアヘッド(one-step ahead)予測因子である;第t日目のアウトプットに対する数値は、アウトプットの過去の数値、並びに被操作変数の現在及び過去の数値から決定される。このモデルは、例えばコントロール戦略により決定されるように、被操作変数に対する規定値と共に、前日に得られたアウトプット予測を使用して将来的なアウトプット値を予測することにより、マルチステップアヘッド予測因子への拡張が可能である。
(式中、
Pは予測ホライズン内の日数である
は、低減次数式モデルに由来する乳酸塩濃度の予測された数値である
rは、所望の参照軌道に対する乳酸塩濃度の数値である
は、予測ホライズン内の各日に対する予測されたアウトプットと参照軌道との間の差異に適用される重み付けである
nmvは、被操作変数の数である
ujは、特定の日における被操作変数jの数値である
は、i番目の予測ホライズン日における被操作変数jに対する、後続する被操作可変数値間の差異に適用される重み付けである
は、被操作変数間のスケールの差異を取り扱うための、j番目の被操作変数に対する倍率である)
(式中、
Pは予測ホライズン内の日数である;
は、低減次数式モデルに由来する乳酸塩濃度の予測値である;rは所望の参照軌道に対する乳酸塩濃度の数値である;
は、予測ホライズン内の各日に対する、予測されたアウトプットと参照軌道との間の差異に適用される重み付けである)を提供するために0に設定され得る。
を1に設定した)。栄養フィード容積は、1.8%~3.6%の間に留まるように限定され、日間の最大変動は3日目~6日目において±1.8%、それ以外は±1.0%に制限された。pHに対する範囲制約を6.7及び7.2に規定し、日にち間pHの最大変動を±0.5に設定した。
](肥満、鼻腔内)、FGLL、アタシセプト、BR3-Fc、BN-003、BA-058、ヒト副甲状腺ホルモン1-34(鼻腔用、骨粗鬆症)、F-18-CCR1、AT-1100(セリアック病/糖尿病)、JPD-003、PTH(7-34)リポソームクリーム(ノバソーム)、デュラマイシン(眼科用、ドライアイ)、CAB-2、CTCE-0214、グリコペグ化エリスロポエチン、EPO-Fc、CNTO-528、AMG-114、JR-013、第VIII因子、アミノカンジン、PN-951、716155、SUN-E7001、TH-0318、BAY-73-7977、テベレリクス(即時放出)、EP-51216、hGH(制御放出、Biosphere)、OGP-I、シフビルチド、TV4710、ALG-889、Org-41259、rhCC10、F-991、チモペンチン(肺疾患)、r(m)CRP、肝臓選択性インスリン、スバリン、L19-IL-2融合タンパク質、エラフィン、NMK-150、ALTU-139、EN-122004、rhTPO、トロンボポエチン受容体作動薬(血小板減少性疾患)、AL-108、AL-208、神経成長因子拮抗薬(疼痛)、SLV-317、CGX-1007、INNO-105、経口テリパラチド(エリゲン)、GEM-OS1、AC-162352、PRX-302、LFn-p24融合ワクチン(テラポア)、EP-1043、肺炎球菌(S.pneumoniae)小児ワクチン、マラリアワクチン、髄膜炎菌(Neisseria meningitidis)B群ワクチン、新生児B群連鎖球菌ワクチン、炭疽病ワクチン、HCVワクチン(gpE1+gpE2+MF-59)、中耳炎治療、HCVワクチン(コア抗原+ISCOMATRIX)、hPTH(1-34)(経皮用、ViaDerm)、768974、SYN-101、PGN-0052、アビスクムミン、BIM-23190、結核ワクチン、マルチエピトープチロシナーゼペプチド、がんワクチン、エンカスチム、APC-8024、GI-5005、ACC-001、TTS-CD3、血管標的化TNF(固形腫瘍)、デスモプレシン(バッカル制御放出型)、オネルセプト、及びTP-9201である。
Claims (23)
- 細胞培養物を増殖させる方法であって、
細胞培養物中の乳酸塩の濃度を決定するステップと、
前記細胞培養物内の少なくとも1つの乳酸塩に影響を及ぼすパラメーターの数値を測定するステップと、
前記乳酸塩濃度、及び前記少なくとも1つの乳酸塩に影響を及ぼすパラメーター測定の前記数値をコントローラーに送付するステップであり、前記コントローラーが、前記少なくとも1つの乳酸塩に影響を及ぼすパラメーターの前記数値に少なくとも部分的に基づき、前記細胞培養物中の乳酸塩の将来的濃度を計算するように構成されている予測モデルを含む、ステップと、
オプティマイザーでシミュレーションを実行して、前記細胞培養物内の乳酸塩に影響を及ぼすパラメーターの操作に基づき、前記乳酸塩の将来的濃度を計算するステップと、
前記コントローラーが、乳酸塩濃度を事前設定された限界内に維持するために、前記細胞培養物中の乳酸塩の計算済みの将来的濃度の前記シミュレーションに基づき、前記細胞培養物内の少なくとも1つの条件を選択的に変化させるステップと、を含み、
前記細胞培養物が、採取される前にインキュベーション期間を有し、前記予測モデルが、インキュベーション期間の終了時の最終乳酸塩濃度を予想する、方法。 - 前記乳酸塩に影響を及ぼすパラメーターが、pH、グルタミン酸濃度、グルコース濃度、アスパラギン濃度、温度、及び/又は栄養フィード速度を含む、請求項1に記載の方法。
- 少なくとも2つの乳酸塩に影響を及ぼすパラメーターが測定され、並びに測定されたデータが前記コントローラーに送付され、及び前記細胞培養物中の乳酸塩の前記将来的濃度を計算するステップで使用される、請求項1に記載の方法。
- 前記少なくとも1つの条件が、前記細胞培養物にフィードされる栄養培地を変化させることにより選択的に変更される、請求項1に記載の方法。
- 前記栄養培地が、炭水化物供給源、アミノ酸供給源、ビタミン、脂質、タンパク質、ペプチド、又はそれらの混合物を含む、請求項4に記載の方法。
- 前記細胞培養物にフィードされる前記栄養培地が、前記細胞培養物に対する前記栄養培地の流速を変化させることにより変更される、請求項4に記載の方法。
- 前記細胞培養物にフィードされる前記栄養培地を変化させるステップに加えて、前記細胞培養物のpHも、乳酸塩濃度を事前設定された限界内に維持するために選択的に変更される、請求項6に記載の方法。
- 前記細胞培養物内の少なくとも1つの条件が、インキュベーション期間の終了時の前記細胞培養物の前記最終乳酸塩濃度が、2g/L未満となるように、インキュベーション期間中に選択的に変更される、請求項1に記載の方法。
- 前記細胞培養物のタイター濃度の増加をもたらす、請求項1に記載の方法。
- 前記細胞培養物が、哺乳動物細胞を含有する、請求項1に記載の方法。
- 前記細胞培養物が、バッチプロセスで12時間~28日間増殖し、次に採取される、請求項1に記載の方法。
- 前記細胞培養物内の前記乳酸塩濃度が、前記コントローラーが前記細胞培養物内の乳酸塩の将来的濃度を計算する前、12時間~4日間において計算される、請求項11に記載の方法。
- 前記バッチプロセスが、前記細胞培養物を採取する前にインキュベーション時間を含み、前記乳酸塩濃度が、前記コントローラーが前記細胞培養物中の乳酸塩の将来的濃度を計算する前に、前記インキュベーション時間の5%~40%において測定される、請求項11に記載の方法。
- 前記乳酸塩濃度が、少なくとも12時間毎に計算され、乳酸塩濃度データのすべてが前記コントローラーにフィードされ、前記コントローラーが、さらなるデータの受領時に、前記細胞培養物内の前記乳酸塩の将来的濃度を反復して計算するように構成されている、請求項11に記載の方法。
- 前記予測モデルが、これまでの参照データに対する乳酸塩濃度の比較に基づく、請求項1に記載の方法。
- 前記乳酸塩の将来的濃度が、規定された参照軌道から予測された乳酸塩濃度の二乗偏差から、前記予測モデルにより計算される、請求項1に記載の方法。
- 前記乳酸塩の予測的濃度が、前記少なくとも1つの乳酸塩に影響を及ぼすパラメーターの変化における二乗偏差に基づき、同様に計算される、請求項16に記載の方法。
- 前記乳酸塩の将来的濃度が、部分的二乗分析、分類木、サポートベクター決定、線形判別分析、又はそれらの混合から選択される1つ又は複数の技術を使用して計算される、請求項1に記載の方法。
- 前記乳酸塩の将来的濃度が、低減次数時間変動式の自己回帰型外因性モデルを使用して、前記コントローラーにより計算される、請求項1に記載の方法。
- 前記予測モデルが、それぞれの日において、予測されたアウトプットと参照された軌道との間の差異に対して重み付けを適用する、請求項16に記載の方法。
- 細胞培養物を増殖させるシステムであって、
細胞培養物を受け入れるための中空内部を規定するバイオリアクターであり、材料を前記中空内部にフィードし、及び/又はそれから取り出すための複数のポートを備えるバイオリアクターと、
栄養培地を前記バイオリアクターの中空内部にフィードするための栄養培地フィードであり、前記バイオリアクターの少なくとも1つのポートと流体連通した状態にある栄養培地フィードと、
前記バイオリアクターに収納された細胞培養物の乳酸塩濃度測定値を受け取るように構成されたコントローラーであり、少なくとも1つの乳酸塩に影響を及ぼすパラメーターの測定値を受け取るように同様に構成されており、前記バイオリアクターに収納された細胞培養物中の乳酸塩の将来的濃度を計算するように構成されている予測モデルを含み、オプティマイザーでシミュレーションを実行して、前記細胞培養物内の栄養培地の操作に基づき、前記乳酸塩濃度のシミュレートされた将来的濃度を計算するように構成されており、予測された乳酸塩濃度の前記シミュレーションに基づき、栄養培地の前記バイオリアクター中への流れを選択的に増加又は減少させて、乳酸塩濃度を事前設定された限界内に維持するために、栄養培地フィードをコントロールするように構成されている、コントローラーと
を備えるシステム。 - 前記少なくとも1つの乳酸塩に影響を及ぼすパラメーターの時間変化を測定するステップを含む、請求項1に記載の方法。
- 前記コントローラーが、閉ループコントロールシステム内で作動し、
前記細胞培養物を含むバイオリアクターと接続したインプット及び/又はアウトプットデバイスに対する調整が、完全に自動化されている、請求項1に記載の方法。
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US11999939B2 (en) | 2024-06-04 |
US20200354666A1 (en) | 2020-11-12 |
KR102589415B1 (ko) | 2023-10-16 |
JP2021503291A (ja) | 2021-02-12 |
EP3714036A1 (en) | 2020-09-30 |
IL274776B1 (en) | 2024-10-01 |
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