JP6101627B2 - 対象粒子を混合物から捕捉する方法 - Google Patents
対象粒子を混合物から捕捉する方法 Download PDFInfo
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- JP6101627B2 JP6101627B2 JP2013535447A JP2013535447A JP6101627B2 JP 6101627 B2 JP6101627 B2 JP 6101627B2 JP 2013535447 A JP2013535447 A JP 2013535447A JP 2013535447 A JP2013535447 A JP 2013535447A JP 6101627 B2 JP6101627 B2 JP 6101627B2
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- IAAKNVCARVEIFS-UHFFFAOYSA-M sodium;4-hydroxynaphthalene-1-sulfonate Chemical compound [Na+].C1=CC=C2C(O)=CC=C(S([O-])(=O)=O)C2=C1 IAAKNVCARVEIFS-UHFFFAOYSA-M 0.000 description 1
- 239000002904 solvent Substances 0.000 description 1
- SFVFIFLLYFPGHH-UHFFFAOYSA-M stearalkonium chloride Chemical compound [Cl-].CCCCCCCCCCCCCCCCCC[N+](C)(C)CC1=CC=CC=C1 SFVFIFLLYFPGHH-UHFFFAOYSA-M 0.000 description 1
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- VLLMWSRANPNYQX-UHFFFAOYSA-N thiadiazole Chemical compound C1=CSN=N1.C1=CSN=N1 VLLMWSRANPNYQX-UHFFFAOYSA-N 0.000 description 1
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- KKSDGJDHHZEWEP-SNAWJCMRSA-N trans-3-coumaric acid Chemical compound OC(=O)\C=C\C1=CC=CC(O)=C1 KKSDGJDHHZEWEP-SNAWJCMRSA-N 0.000 description 1
- NGSWKAQJJWESNS-ZZXKWVIFSA-N trans-4-coumaric acid Chemical compound OC(=O)\C=C\C1=CC=C(O)C=C1 NGSWKAQJJWESNS-ZZXKWVIFSA-N 0.000 description 1
- VZCYOOQTPOCHFL-UHFFFAOYSA-N trans-butenedioic acid Natural products OC(=O)C=CC(O)=O VZCYOOQTPOCHFL-UHFFFAOYSA-N 0.000 description 1
- 230000014616 translation Effects 0.000 description 1
- 238000004627 transmission electron microscopy Methods 0.000 description 1
- 239000013638 trimer Substances 0.000 description 1
- GPRLSGONYQIRFK-MNYXATJNSA-N triton Chemical compound [3H+] GPRLSGONYQIRFK-MNYXATJNSA-N 0.000 description 1
- 229960003732 tyramine Drugs 0.000 description 1
- 125000002948 undecyl group Chemical group [H]C([*])([H])C([H])([H])C([H])([H])C([H])([H])C([H])([H])C([H])([H])C([H])([H])C([H])([H])C([H])([H])C([H])([H])C([H])([H])[H] 0.000 description 1
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- 229910052727 yttrium Inorganic materials 0.000 description 1
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- 239000002888 zwitterionic surfactant Substances 0.000 description 1
- DGVVWUTYPXICAM-UHFFFAOYSA-N β‐Mercaptoethanol Chemical compound OCCS DGVVWUTYPXICAM-UHFFFAOYSA-N 0.000 description 1
Classifications
-
- B—PERFORMING OPERATIONS; TRANSPORTING
- B01—PHYSICAL OR CHEMICAL PROCESSES OR APPARATUS IN GENERAL
- B01D—SEPARATION
- B01D15/00—Separating processes involving the treatment of liquids with solid sorbents; Apparatus therefor
- B01D15/08—Selective adsorption, e.g. chromatography
- B01D15/10—Selective adsorption, e.g. chromatography characterised by constructional or operational features
- B01D15/18—Selective adsorption, e.g. chromatography characterised by constructional or operational features relating to flow patterns
-
- C—CHEMISTRY; METALLURGY
- C07—ORGANIC CHEMISTRY
- C07K—PEPTIDES
- C07K1/00—General methods for the preparation of peptides, i.e. processes for the organic chemical preparation of peptides or proteins of any length
- C07K1/14—Extraction; Separation; Purification
- C07K1/16—Extraction; Separation; Purification by chromatography
- C07K1/18—Ion-exchange chromatography
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K39/00—Medicinal preparations containing antigens or antibodies
-
- B—PERFORMING OPERATIONS; TRANSPORTING
- B01—PHYSICAL OR CHEMICAL PROCESSES OR APPARATUS IN GENERAL
- B01D—SEPARATION
- B01D15/00—Separating processes involving the treatment of liquids with solid sorbents; Apparatus therefor
- B01D15/08—Selective adsorption, e.g. chromatography
- B01D15/10—Selective adsorption, e.g. chromatography characterised by constructional or operational features
- B01D15/18—Selective adsorption, e.g. chromatography characterised by constructional or operational features relating to flow patterns
- B01D15/1807—Selective adsorption, e.g. chromatography characterised by constructional or operational features relating to flow patterns using counter-currents, e.g. fluidised beds
-
- B—PERFORMING OPERATIONS; TRANSPORTING
- B01—PHYSICAL OR CHEMICAL PROCESSES OR APPARATUS IN GENERAL
- B01J—CHEMICAL OR PHYSICAL PROCESSES, e.g. CATALYSIS OR COLLOID CHEMISTRY; THEIR RELEVANT APPARATUS
- B01J20/00—Solid sorbent compositions or filter aid compositions; Sorbents for chromatography; Processes for preparing, regenerating or reactivating thereof
- B01J20/281—Sorbents specially adapted for preparative, analytical or investigative chromatography
-
- C—CHEMISTRY; METALLURGY
- C07—ORGANIC CHEMISTRY
- C07K—PEPTIDES
- C07K14/00—Peptides having more than 20 amino acids; Gastrins; Somatostatins; Melanotropins; Derivatives thereof
- C07K14/415—Peptides having more than 20 amino acids; Gastrins; Somatostatins; Melanotropins; Derivatives thereof from plants
- C07K14/42—Lectins, e.g. concanavalin, phytohaemagglutinin
-
- B—PERFORMING OPERATIONS; TRANSPORTING
- B01—PHYSICAL OR CHEMICAL PROCESSES OR APPARATUS IN GENERAL
- B01D—SEPARATION
- B01D15/00—Separating processes involving the treatment of liquids with solid sorbents; Apparatus therefor
- B01D15/08—Selective adsorption, e.g. chromatography
- B01D15/26—Selective adsorption, e.g. chromatography characterised by the separation mechanism
- B01D15/36—Selective adsorption, e.g. chromatography characterised by the separation mechanism involving ionic interaction
- B01D15/361—Ion-exchange
- B01D15/363—Anion-exchange
-
- C—CHEMISTRY; METALLURGY
- C07—ORGANIC CHEMISTRY
- C07K—PEPTIDES
- C07K1/00—General methods for the preparation of peptides, i.e. processes for the organic chemical preparation of peptides or proteins of any length
- C07K1/14—Extraction; Separation; Purification
Landscapes
- Chemical & Material Sciences (AREA)
- Organic Chemistry (AREA)
- Health & Medical Sciences (AREA)
- Life Sciences & Earth Sciences (AREA)
- Analytical Chemistry (AREA)
- General Health & Medical Sciences (AREA)
- Medicinal Chemistry (AREA)
- Biophysics (AREA)
- Genetics & Genomics (AREA)
- Biochemistry (AREA)
- Molecular Biology (AREA)
- Proteomics, Peptides & Aminoacids (AREA)
- Chemical Kinetics & Catalysis (AREA)
- Botany (AREA)
- Gastroenterology & Hepatology (AREA)
- Microbiology (AREA)
- Immunology (AREA)
- Mycology (AREA)
- Pharmacology & Pharmacy (AREA)
- Epidemiology (AREA)
- Animal Behavior & Ethology (AREA)
- Public Health (AREA)
- Veterinary Medicine (AREA)
- Solid-Sorbent Or Filter-Aiding Compositions (AREA)
- Micro-Organisms Or Cultivation Processes Thereof (AREA)
- Peptides Or Proteins (AREA)
- Treatment Of Liquids With Adsorbents In General (AREA)
Description
(i)吸着剤の密度が約2.5g/ml〜3.5g/mlの間であり、平均吸着剤粒子サイズが約50である、ジエチルアミノエチルセルロースデキストランを含む吸着剤の拡張ベッドをカラム中に提供する工程と;
(ii)約pH6.0〜8.0範囲のpHで樹脂材料を平衡化する工程と;
(iii)タバコ植物材料を含む混合物であって、約2mS/cm〜3mS/cmの間の伝導率を有する前記混合物を提供する工程と;
(iv)カラム中の拡張ベッドの拡張度が約2で、約2.5cm/分のフロー率で、混合物をカラム上にロードしてウイルス様粒子を結合させる工程と;
(v)約pH6.0〜8.0の範囲のpHを有するバッファーを使用してロードしたカラムを洗浄する工程と;
(vi)1つまたは複数の画分でカラムから対象となる結合粒子を溶出する工程と
を含む、対象となるウイルス様粒子を混合物から捕捉するために提供される。
定義
インフルエンザウイルス様粒子は、D’Aoust,M.−A.,Lavoie,P.−O.,Couture,M.M.−J.,Trepanier,S.,Guay,J.−M.,Dargis,M.,Mongrand,S.,Landry,N.,Ward,B.J.,Vezina,L.−P.Influenza virus−like particles produced by transient expression in Nicotiana benthamiana induce a protective immune response against a lethal viral challenge.Plant Biotechnology Journal 6 (2008) 930−940中で記載されるようなNicotiana benthamiana植物において生産される。
赤血球凝集はWO2004/098533中で記載されるような赤血球によるインキュベーションによって測定される。赤血球凝集素タンパク質は標準的方法を使用してELISAによって検出される。赤血球凝集素H5タンパク質の検出については、Immunetechからのウサギ抗H5抗体(カタログIT−003−005V)およびJacksonからのホースラディシュペルオキシダーゼ標識二次抗体(カタログ11−035−046)を使用する。
実施例1中におよびD’Aoust et al.(2008年、前出)によって記載されるように、インフィルトレーションされたN.benthamiana植物を収穫前に6日間温室でインキュベーションする。葉の収穫に際して、バイオマスを暗所で4℃で一晩維持し、コーン上に少なくとも45psiの圧力を使用する15〜20kg/時間のねじプレス(Vincent CP−4)を使用してホモジナイズする。緑色ジュース抽出物を回収し、メタ重亜硫酸ナトリウムを10mMの最終濃度に緑色ジュースに添加する。伝導率を測定し、必要に応じて抽出物を捕捉実験での使用前に水で希釈する。
抽出物。赤血球凝集素H5ウイルス様粒子(D’Aoust et al.(2008)、前出)を含むN.benthamiana緑色ジュース抽出物を実施例3中に記載されるようなねじプレスを使用して得た。緑色ジュース抽出物を純水中で5倍に希釈し(最終pH7)、伝導率を測定した。伝導率は5.6mS/cmであった。以下の異なる4つのリガンドを結合について試験した。ジエチルアミノエチル(DEAE)イオン交換体、デキストランベースのDEAEイオン交換体、デキストランベースのクロロメチル−2−ベンズイミダゾールおよびデキストランベースの1−(2−クロロエチル)塩酸ピペリジンリガンド。様々な吸着剤対使用されるサンプルの比率は1:5であった。静的モードにおけるバッチインキュベーション後に、上清を回収し、赤血球凝集を実施例2の手順に従って測定した。
実施例1および3中の記述に従って得られるような、赤血球凝集素H5ウイルス様粒子を含む緑色ジュース抽出物の伝導率の、静的結合モードにおけるDEAEリガンドへのウイルス様粒子の結合に対する効果を測定して、どのくらい高い伝導率でも緑色ジュース抽出物中のウイルス様粒子がDEAEイオン交換体に結合できるかを調べる。
2工程プロセス。抽出物および特に緑色の原因となる抽出物中の任意の材料が、大部分のリガンドに粘着し、カラムの目詰まりおよび閉塞を引き起こすので、実施例3中に記載される方法に従って得られた緑色植物ジュース抽出物は、前捕捉を利用する。この目的のために、植物に由来するウイルス様粒子のための2工程の捕捉プロセスを設定した。プロセスは以下のものを含んでいた。
工程2、pH7で50μmの平均粒子径を備えたDEAEイオン交換体を使用して、拡張ベッド吸着モードにおいて、工程1の上清からのウイルス様粒子を結合させること。
実施例6中に記載されるような充填ベッドモードにおけるDEAEイオン交換体におよび続いてPEIイオン交換体による前捕捉による、実施例3中に記載されていた方法に従って得られた緑色植物ジュース抽出物中に存在するウイルス様粒子の結合および溶出を、50mMトリス/HCl、pH8.5を含む溶出バッファーに1M NaCl、0.8M NaCl、0.6M NaCl、0.4M NaCl、0.2M NaClまたは0.1M NaClを添加したものを使用して試験した。抽出物のローディング前に、緑色植物ジュース抽出物を、脱イオン水中で5倍に希釈し、pHをpH7に調節した。伝導率は5.1mS/cmであった。実施例6の工程1で記載されるように、濾過された緑色植物ジュース抽出物を、バッチインキュベーションモードにおいて、15:1の抽出物対リガンドの比率で、150μmの平均粒子径のPEIイオン交換体により最初に30分間インキュベーションした。使用されるDEAEイオン交換体は50μmの平均粒子径を有し、上清対吸着剤の比率は5:1であった。実施例2中に記載されるように赤血球凝集素タンパク質の量を測定した。素通りおよび洗浄物、DEAEイオン交換体に結合したもの、ならびに溶出物中の赤血球凝集素の相対量を、表4中に示す。この実験において、0.4、0.6、0.8または1.0MのNaClを含む溶出バッファーの使用は、赤血球凝集アッセイを使用して測定されるウイルス様粒子の類似した捕捉をもたらした。
緑色植物ジュース抽出物中に存在し、実施例3中に記載される方法に従って得られたウイルス様粒子の結合を、拡張ベッド吸着モードの1工程の結合プロセスにおいて、228cm/時間および450cm/時間のフロー率で測定した。5ベッド体積の0.5Mトリス/HCl、pH7バッファーおよび10ベッド体積の10mMトリス、pH7を使用して平衡化したDEAEイオン交換体を使用した。使用されるDEAEイオン交換体は50μmの平均粒子径を有していた。吸着剤対緑色植物ジュース抽出物の比率は常に1:20だった。ベッド高は50cmであり、カラムは39.3mLの吸着剤DEAEイオン交換体を含んでいた。緑色植物ジュース抽出物を脱イオン水中で5倍に希釈し、ローディング前に伝導率を測定した。洗浄バッファーは10mMトリス、pH7.0、および溶出バッファーは50mMトリス/HCl、1M NaCl、pH9.0であった。
前処理された緑色植物ジュース抽出物中に存在し、実施例3中に記載される方法に従って得られたウイルス様粒子の結合を、本質的に実施例8中に記載されるように、拡張ベッド吸着モードの1工程の結合プロセスにおいて、450cm/時間のフロー率で測定した。すべての実験条件は、初期抽出物の伝導率が希釈後に5.36mS/cmだったことおよび抽出物が拡張ベッド吸着モードにおける1工程の結合プロセスに適用する前に酸沈殿および濾過されたこと以外は、実施例8において使用されたものに類似していた。独立した2つの実験を実行した。両方の実験についての赤血球凝集アッセイの結果を、素通り画分1〜3、組み合わせた素通りおよび洗浄物画分、および組み合わせた素通りおよび洗浄物、および溶出物について表7中に示す。
抽出物。記載される(D’Aoust et al.(2008)、前出)ように、7,500のN.benthamiana植物をインフィルトレーションして、赤血球凝集素H5ウイルス様粒子を含む250kgの粗製バイオマスを生成する。インフィルトレーション後に、植物を収穫前に6日間温室でインキュベーションする。本質的に実施例3中に記載されるように、葉の収穫に際して、バイオマスを暗所で4℃で一晩維持し、コーン上に少なくとも45psiの圧力を使用する15〜20kg/時間のねじプレス(Vincent CP−4)を使用してホモジナイズする。緑色ジュースを回収し、メタ重亜硫酸ナトリウムを10mMの最終濃度で緑色ジュースに添加する。伝導率を測定し、使用前に抽出物を約10の最終伝導率に水により希釈する。抽出物を室温で維持する。希釈後の全体積は約1250Lの抽出物である。
溶出後に、樹脂を4カラム体積の0.5M NaOH続いて4カラム体積の蒸留水による洗浄によって浄化および再生する。再平衡を10mMトリス−Clバッファー、pH7中で行う。
アグロ−インフィルトレーションの6日後に収穫されたH5−VLPを発現するN.benthamiana植物を、4℃で一晩暗所で維持する。葉および茎を含む植物バイオマスを、ねじプレス(Green Star Corrupad、GS、Korea Co.1000)を使用してホモジナイズする。MBSを10mMの最終濃度で添加してサンプル酸化を回避する。ねじプレスから得られた粗製ジュースを、20%に希釈した酢酸を使用してpH5.3に迅速に調節する。抽出物は部分的清澄化のために撹拌せずに室温で20〜30分間放置する。濾過は、高さ3mmのCelpure P300(MBS 10mM中の10%Celpure P300スラリー)により前コートされたWhatmanの濾紙を通して実行する。真空を508mbarで維持した。Celpure P300(10%)を抽出物に添加し、1分間混合する。最終的に、抽出物−celpureスラリーを穏やかに添加して、抽出物を小分けにして濾過する(濾過ケーキにわたって2cm以下の液相)。可能性のある4つの吸収剤を選択し、それらの結合条件をバッチモードにおける静的結合条件下で最適化する。表9中の結果は、イオン交換体吸着剤の結合能力が以下の2つの因子によって影響を受けることを示す。(i)低イオン強度(2.3mS/cm以下)および従って(ii)吸着剤の単位質量あたりの植物抽出物の高体積(20mlの抽出物に対して1gの吸着剤)。
Claims (13)
- ウイルス様粒子を含む植物の破壊細胞の抽出物を含む混合物から対象となるウイルス様粒子を捕捉する方法であり;
(i)樹脂を含む吸着剤の拡張ベッドを提供する工程;
(ii)約6.0〜8.0の範囲のpHで前記樹脂を平衡化する工程;
(iii)約2cm/分〜8cm/分の間の範囲のフロー率で前記混合物を前記吸着剤の拡張ベッド上にロードしてウイルス様粒子を結合させる工程;
(iv)任意で前記吸着剤を洗浄する工程;および
(v)任意で前記吸着剤からウイルス様粒子を溶出する工程、を含み、
吸着剤に接触する前の混合物の伝導率が2mS/cm〜10mS/cmの間であり、
溶出工程(v)の吸着剤の拡張ベッドの拡張度が約0.5〜5の範囲であり、
「約」という用語は、与えられた数の値または範囲の文脈において、与えられた値または範囲の20%内であり、及び
前記抽出物は前処理されておらず、1工程の結合及び溶出プロセスにおいて、対象となる粒子を単離することができる、
前記方法。 - 前記吸着剤がビーズの形態で提供される、請求項1に記載の方法。
- 前記吸着剤がカラム内に含まれる、請求項1または2に記載の方法。
- 前記ウイルス様粒子が、インフルエンザウイルスのタンパク質を含む、請求項1〜3のいずれか一項に記載の方法。
- 前記ウイルス様粒子が、赤血球凝集素;H1、H2、H3、H4、H5、H6、H7、H8、H9、H10、H11、H12、H13、H14、H15もしくはH16またはそれらの2つ以上の組み合わせからなる群から選択される赤血球凝集素サブタイプを含む、請求項1〜4のいずれか一項に記載の方法。
- 前記混合物が、タンパク質を植物中で一過性に発現する核酸分子によりインフィルトレーションされたタバコ植物の破壊細胞を含み;前記タンパク質がウイルス様粒子中に存在し;前記混合物が植物の地上部であるかまたは植物の地上部に由来する、請求項1〜5のいずれか一項に記載の方法。
- 前記ビーズが、プラスチック、メタクリレート、陰イオン交換体、ジエチルアミノエチル、陽イオン交換体、多糖、シリカ、ポリ(スチレンジビニル)ベンゼン、ポリアクリルアミド、セラミックおよびその誘導体またはその2つ以上の組み合わせからなる群から選択される材料を含むか、それらからなるか、または本質的になり;前記ビーズが、セルロース、アガロースおよびデキストランならびにその誘導体またはその2つ以上の組み合わせからなる群から選択される多糖を含むか、それらからなるか、または本質的になる、請求項2〜6のいずれか一項に記載の方法。
- 前記ビーズが、石英、シリカ、Nd−Fe−B合金、ステンレス鋼、酸化ジルコニウム、ジルコニア、金属ケイ酸塩、金属硼ケイ酸塩、セラミック(二硼化チタン、炭化チタン、二硼化ジルコニウム、炭化ジルコニウム、炭化タングステン、炭化ケイ素、窒化アルミニウム、窒化ケイ素、窒化チタン、酸化イットリウム、シリコン金属粉末および二ケイ化モリブデンを含む);酸化マグネシウム、酸化アルミニウム、酸化チタン、酸化バナジウム、酸化クロミウム、酸化ジルコニウム、酸化ハフニウム、酸化マンガン、酸化鉄、酸化コバルト、酸化ニッケル、酸化銅および酸化銀を含む、金属酸化物および硫化物;非金属酸化物;硫酸バリウムを含む金属塩;タングステン、ジルコニウム、チタン、ハフニウム、バナジウム、クロミウム、マンガン、鉄、コバルト、ニッケル、インジウム、銅、銀、金、パラジウム、プラチナ、ルテニウム、オスミウム、ロジウムおよびイリジウムを含む、金属元素、ならびに前記金属元素およびその誘導体またはその2つ以上の組み合わせの間に形成された合金等の金属元素の合金からなる群から選択される材料を含むか、それらからなるか、または本質的になる不活性コアを含む、請求項7に記載の方法。
- 対象となる粒子を結合する前記ビーズのサイズが約25μm〜100μmの間であり;「約」という用語は、与えられた数の値または範囲の文脈において、与えられた値または範囲の20%内である、請求項2〜8のいずれか一項に記載の方法。
- 前記吸着剤がリガンドを含むか、それからなるか、または本質的になり;前記リガンドが、(i)ベンジルアミンまたはその誘導体;(ii)アルキルアミンもしくはその誘導体;(iii)アルケニルアミンもしくはその誘導体;(iv)アルキニルアミンもしくはその誘導体;(v)アルコキシルアミンもしくはその誘導体;および(vi)単環式もしくは二環式の芳香族もしくはヘテロ芳香族のモイエティもしくはその誘導体のアミン、またはその2つ以上の組み合わせからなる群から選択されるリガンドを含むか、それらからなるか、または本質的になるか;または前記リガンドが陰イオン交換樹脂を含むか、それからなるか、または本質的になり;前記陰イオン交換樹脂が、ジエチルアミノエチルセルロースベースのイオン交換樹脂を含むか、それからなるか、もしくは本質的になるか、またはジエチルアミノエチルセルロースデキストランベースのイオン交換樹脂を含むか、それからなるか、もしくは本質的になる、請求項1〜9のいずれか一項に記載の方法。
- (i)吸着剤の密度が約2.5g/ml〜3.5g/mlの間であり、平均吸着剤粒子サイズが約50μmである、ジエチルアミノエチルセルロースデキストランを含む吸着剤の拡張ベッドをカラム中に提供する工程と;
(ii)約pH6.0〜8.0範囲のpHで樹脂材料を平衡化する工程と;
(iii)タバコ植物材料を含む混合物であって、約2mS/cm〜3mS/cmの間の伝導率を有する前記混合物を提供する工程と;
(iv)カラム中の吸着剤の拡張ベッドの拡張度が約2で、約2.5cm/分のフロー率で、混合物をカラム上にロードしてウイルス様粒子を結合させる工程と;
(v)約pH6.0〜8.0の範囲のpHを有するバッファーを使用してロードしたカラムを洗浄する工程と;
(vi)カラムから対象となる結合粒子を溶出して1つまたは複数の画分にする工程と
を含み、
「約」という用語は、与えられた数の値または範囲の文脈において、与えられた値または範囲の20%内である、請求項1〜10のいずれか一項に記載の方法。 - 工程(iii)が、(a)ウイルス様粒子中に存在するタンパク質を一過性に発現する全体のタバコ植物を提供することと;(b)タバコ植物の葉および茎の細胞を破壊してタバコ植物材料を産生することと;(c)バッファーまたは脱イオン水による希釈によって混合物のpHおよび伝導率を合わせることとを含む、請求項11に記載の方法。
- 工程(iv)の前に、工程(iii)の混合物を濾過することおよび工程(iii)の濾過された混合物をビーズのサイズが約150μmであるポリエチレンイミンを含む吸着剤の第1の拡張ベッドと接触させて所望されない物質を混合物から除去することを含む工程をさらに含み、「約」という用語は、与えられた数の値の文脈において、与えられた値の20%内である、請求項11に記載の方法。
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KR20230056661A (ko) | 2020-08-28 | 2023-04-27 | 가부시키가이샤 가네카 | 유용 물질의 정제 방법 |
GB202109610D0 (en) * | 2021-07-02 | 2021-08-18 | Cytiva Bioprocess R & D Ab | Chromatography medium for use in purification of enveloped virus particles or exosomes |
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LU73094A1 (ja) * | 1975-07-29 | 1977-03-24 | ||
FR2422720A2 (fr) * | 1977-04-26 | 1979-11-09 | Elf Aquitaine | Separation et purification de proteines par chromatographie |
WO1991013555A1 (en) | 1990-03-07 | 1991-09-19 | Engelhard Corporation | Animal feed additive and method for inactivating mycotoxins present in animal feeds |
SE9101149D0 (sv) * | 1991-04-17 | 1991-04-17 | Pharmacia Lkb Biotech | Beads for down stream processing |
FR2679448B1 (fr) | 1991-07-23 | 1993-10-15 | Oreal | Melange azetrope de thioglycolate d'hydroxy-2 propyle et de thioglycolate d'hydroxy-2 methyl-1 ethyle, son procede d'obtention et son utilisation dans un procede de deformation permanente des cheveux. |
DE69933677T2 (de) * | 1998-08-11 | 2007-08-23 | Biosource Technologies, Inc., Vacaville | Verfahren zur gewinnung von proteinen aus der interstitiellen flüssigkeit von pflanzen |
DE60029195T2 (de) * | 1999-02-22 | 2007-06-28 | Transgene S.A. | Verfahren zur Gewinnung von purifizierter Virenzusammensetzung |
ATE365463T1 (de) | 2003-03-21 | 2007-07-15 | Upfront Chromatography As | Verfahren für volumen mit hohem durchsatz bei der fraktionierung von biomolekülen durch chromatographische systeme |
BRPI0410340A (pt) | 2003-05-05 | 2006-05-30 | Dow Agrosciences Llc | vetores e células para preparar composições imunoprotetoras derivadas de plantas transgênicas |
AU2004269199B2 (en) * | 2003-08-27 | 2009-06-25 | Orf Liftaekni Hf. | A process for proteolytic cleavage and purification of recombinant proteins produced in plants |
EA013504B1 (ru) * | 2005-04-11 | 2010-06-30 | Медарекс, Инк. | Способ очистки белков |
EP1736538A1 (en) * | 2005-06-21 | 2006-12-27 | Cytos Biotechnology AG | Process for the preparative purification of virus-like-particles (VLPs) |
DE102006002249B4 (de) * | 2006-01-17 | 2010-12-09 | Fraunhofer-Gesellschaft zur Förderung der angewandten Forschung e.V. | Verfahren zur Gewinnung einer pflanzlichen Proteinfraktion, insbesondere zur Herstellung von pflanzlichem Speiseeis |
BRPI0806601A2 (pt) * | 2007-01-15 | 2011-09-06 | Upfront Chromatography As | produção de biocombustìvel e proteìna a partir de um material bruto |
ATE515301T1 (de) * | 2007-05-15 | 2011-07-15 | Cooperatie Avebe U A | Verfahren und vorrichtung für kontinuierliche chromatografische trennung |
CA2615372A1 (en) * | 2007-07-13 | 2009-01-13 | Marc-Andre D'aoust | Influenza virus-like particles (vlps) comprising hemagglutinin |
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KR20130122947A (ko) | 2013-11-11 |
CN103228327A (zh) | 2013-07-31 |
RU2013124062A (ru) | 2014-12-10 |
CN103228327B (zh) | 2016-01-27 |
WO2012055986A1 (en) | 2012-05-03 |
WO2012055986A9 (en) | 2012-06-28 |
EP2632562A1 (en) | 2013-09-04 |
EP2632562B1 (en) | 2020-08-05 |
US20130317197A1 (en) | 2013-11-28 |
JP2013544509A (ja) | 2013-12-19 |
RU2606609C2 (ru) | 2017-01-10 |
BR112013010385A2 (pt) | 2016-08-02 |
US9056894B2 (en) | 2015-06-16 |
CA2816190A1 (en) | 2012-05-03 |
SG190031A1 (en) | 2013-06-28 |
JP2017055766A (ja) | 2017-03-23 |
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