Process for separating alpha-1-proteinase inhibitor from...

C - Chemistry – Metallurgy – 07 – K

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C07K 14/81 (2006.01) C07K 1/14 (2006.01) C07K 1/36 (2006.01)

Patent

CA 2259499

The present invention is directed to a process for purifying alpha-1-PI. The process comprises providing an impure protein fraction which comprises alpha-1-PI. The impure protein fraction is suspended in an aqueous solution at pH 6. Insoluble proteins are recovered and resuspended in aqueous solution at pH 8,5. PEG is added to precipitate .alpha.-2 proteins. To the PEG supernatant precipitation, which comprises alpha-1-PI, is added ZnCl2 to precipitate crude alpha-1-PI. The crude alpha-1-PI is resolubilized and apllied to an anion-exchange medium. A fraction comprising alpha-1-PI is recovered from the anion-exchange medium. Alpha-1-PI purified by the process has a specific activity about 1.0 units/OD280.

La présente invention a trait à un procédé de purification de l'inhibiteur de protéinase alpha -1 (alpha-1-PI). Le procédé comprend l'obtention d'une fraction de protéine impure comportant de l'alpha-1-PI. La fraction de protéine impure est suspendue dans une solution aqueuse à pH 6. Les protéines insolubles sont récupérées et resuspendues dans une solution aqueuse à pH 8,5. Du PEG est ajouté pour précipiter les protéines alpha -2. On ajoute audit précipité, qui contient de l'alpha-1-PI, du ZnCl2 pour précipiter l'alpha-1-PI brute. Celle-ci est resolubilisée et appliquée à un support échangeur d'anions. Une fraction comportant de l'alpha-1-PI est récupérée du support échangeur d'anions. L'alpha-1-PI purifiée à l'aide de ce procédé possède une activité spécifique d'environ 1 unité/OD280.

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