PROTEIN PURIFICATION

• Main Authors: MCDONALD PAUL J, EMERY JEFFERSON C, O'LEARY RHONA
• Format: Patent
• Language: English; Japanese
• Published: 23.07.2015
• Subjects: CHEMISTRY; METALLURGY; ORGANIC CHEMISTRY; PEPTIDES; PERFORMING OPERATIONS; PHYSICAL OR CHEMICAL PROCESSES OR APPARATUS IN GENERAL; SEPARATION; TRANSPORTING

PROBLEM TO BE SOLVED: To provide a method for purifying a polypeptide (e.g. an antibody).SOLUTION: A method for purifying a polypeptide from a composition comprising the polypeptide and contaminants comprises the sequential steps of: (a) loading the composition onto an ion exchange resin with an equilibration buffer having a first salt concentration; (b) washing the ion exchange resin with a wash buffer until a predetermined protein concentration is measured in the flowthrough, where the salt concentration of the wash buffer increases from an initial, second salt concentration that is greater than the salt concentration of the equilibration buffer, to a final, third salt concentration; (c) passing a fixed volume of wash buffer at the final, third salt concentration over the cation exchange resin; and (d) eluting the polypeptide from the ion exchange resin with an elution buffer that has a salt concentration that is greater than the final salt concentration of the wash buffer. 【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