PROTEIN PURIFICATION

• Main Authors: MCDONALD PAUL J, EMERY JEFFERSON C, O'LEARY RHONA
• Format: Patent
• Language: English; Japanese
• Published: 22.03.2019
• Subjects: CHEMISTRY; METALLURGY; ORGANIC CHEMISTRY; PEPTIDES; PERFORMING OPERATIONS; PHYSICAL OR CHEMICAL PROCESSES OR APPARATUS IN GENERAL; SEPARATION; TRANSPORTING

To provide methods for purifying polypeptides such as antibodies.SOLUTION: The invention provides a method for purifying a polypeptide from a composition comprising the polypeptide and contaminants comprising the following series of steps: (a) loading the composition to an ion exchange resin with an equilibration buffer of a first salt concentration; (b) washing the ion exchange resin with a washing buffer until a certain protein concentration is detected in the flow-through fraction where the washing buffer has a salt concentration that increases from a second salt concentration higher than that of the equilibration buffer to a final third salt concentration;(c) running a certain volume of the washing buffer in the final third salt concentration through the cation exchange resin; and (d) eluting the polypeptide from the cation exchange resin using an elution buffer of a salt concentration higher than the final salt concentration of the washing buffer.SELECTED DRAWING: None 【課題】ポリペプチド（例えば、抗体）を精製する方法を提供すること。【解決手段】ポリペプチドおよび夾雑物を含む組成物から該ポリペプチドを精製するための方法であって、該方法は、以下の一連の工程：（ａ）該組成物を、第１塩濃度を有する平衡化緩衝液とともにイオン交換樹脂にローディングする工程；（ｂ）該イオン交換樹脂を、素通り画分中に所定のタンパク質濃度が測定されるまで、洗浄緩衝液で洗浄する工程であって、該洗浄緩衝液の塩濃度は、該平衡化緩衝液の塩濃度よりも高い最初の第２塩濃度から、最終的な第３塩濃度へと上昇する、工程；（ｃ）該最終的な第３塩濃度において、一定体積の洗浄緩衝液を、陽イオン交換樹脂に通過させる工程；および（ｄ）該ポリペプチドを、該洗浄緩衝液の最終塩濃度よりも高い塩濃度を有する溶出緩衝液を用いて、該イオン交換樹脂から溶出させる工程を包含する、方法。【選択図】なし