有哪些信誉好的足球投注网站urate oxidase purification by salting-in crystallization towards an alternative to chromatography尿酸盐氧化酶salting-in结晶对替代色谱纯化.pdfVIP
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urate oxidase purification by salting-in crystallization towards an alternative to chromatography尿酸盐氧化酶salting-in结晶对替代色谱纯化
Urate Oxidase Purification by Salting-in Crystallization: Towards an Alternative to Chromatography 1 ´ 1 2 3 3 Marion Giffard , Natalie Ferte , Franc¸ois Ragot , Mohamed El Hajji , Bertrand Castro , Franc¸oise ´ 1* Bonnete ´ 1 Centre Interdisciplinaire de Nanoscience de Marseille, UPR3118 CNRS, Aix-Marseille Universite, Marseille, France, 2 Biotechnology Department, Sanofi-Aventis, Aramon, France, 3 Analytical Sciences Department, Sanofi-Aventis Research and Development, Montpellier, France Abstract Background: Rasburicase (FasturtecH or ElitekH, Sanofi-Aventis), the recombinant form of urate oxidase from Aspergillus flavus, is a therapeutic enzyme used to prevent or decrease the high levels of uric acid in blood that can occur as a result of chemotherapy. It is produced by Sanofi-Aventis and currently purified via several standard steps of chromatography. This work explores the feasibility of replacing one or more chromatography steps in the downstream process by a crystallization step. It compares the efficacy of two crystallization techniques that have proven successful on pure urate oxidase, testing them on impure urate oxidase solutions. Methodology/Principal Findings: Here we investigate the possibility of purifying urate oxidase directly by crystallization from the fermentation broth. Based on attractive interaction potentials which are known to drive urate oxidase crystallization, two crystallization routes are compared: a) by increased polymer concentration, which induces a depletion attraction and b) by decreased salt concentration, which induces attractive interactions via a salting-in
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