Implementation of a crystallization step into the purification process of a recombinant protein

Identification of crystallization conditions of new proteins is still regarded as a tedious trial-and-error work, especially when the crystallization step has to meet the requirements of a given purification process. The traditional screening kit method and a multifactorial approach were compared ag...

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Veröffentlicht in:Protein expression and purification 2005, Vol.39 (1), p.43-53
Hauptverfasser: Peters, Jörg, Minuth, Torsten, Schröder, Werner
Format: Artikel
Sprache:eng
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Zusammenfassung:Identification of crystallization conditions of new proteins is still regarded as a tedious trial-and-error work, especially when the crystallization step has to meet the requirements of a given purification process. The traditional screening kit method and a multifactorial approach were compared against each other with regard to their ability to find new crystallization conditions that are compatible to the purification process of a recombinant aprotinin variant. Overall, the multifactorial approach turned out to be 10-fold more efficient. The new crystallization conditions were scaled up and implemented into the purification process as a bulk storage step. The aprotinin variant derived from this process was fully characterized biochemically.
ISSN:1046-5928
1096-0279
DOI:10.1016/j.pep.2004.09.011