Ice-shell purification of ice-binding proteins

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• 作者：Craig J. Marshall^a ; ^{craig.marshall@otago.ac.nz" class="auth_mail" title="E-mail the corresponding author} ; Koli Basu^b ; Peter L. Davies^b
• 关键词：Antifreeze protein ; Ice affinity ; Ice shell ; Ice finger
• 刊名：Cryobiology
• 出版年：2016
• 出版时间：June 2016
• 年：2016
• 卷：72
• 期：3
• 页码：258-263
• 全文大小：1056 K

文摘

Ice-affinity purification is a simple and efficient method of purifying to homogeneity both natural and recombinant ice-binding proteins. The purification involves the incorporation of ice-binding proteins into slowly-growing ice and the exclusion of other proteins and solutes. In previous approaches, the ice was grown around a hollow brass finger through which coolant was circulated. We describe here an easily-constructed apparatus that employs ice affinity purification that not only shortens the time for purification from 1–2 days to 1–2 h, but also enhances yield and purity. In this apparatus, the surface area for the separation was increased by extracting the ice-binding proteins into an ice-shell formed inside a rotating round-bottom flask partially submerged in a sub-zero bath. In principle, any ice-binding compound can be recovered from liquid solution, and the method is readily scalable.