title: Ice-shell purification of ice-binding proteins.
abstract: 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.
authors.affiliation: Department of Biochemistry, and Genetics Otago, University of Otago, PO Box 56, Dunedin, 9054, New Zealand. Electronic address: craig.marshall@otago.ac.nz.
authors.forename: Craig J
authors.initials: CJ
authors.lastname: Marshall
authors.affiliation: Department of Biomedical and Molecular Sciences, Queen's University, Kingston, Ontario, K7L 3N6, Canada.
authors.forename: Koli
authors.initials: K
authors.lastname: Basu
authors.affiliation: Department of Biomedical and Molecular Sciences, Queen's University, Kingston, Ontario, K7L 3N6, Canada.
authors.forename: Peter L
authors.initials: PL
authors.lastname: Davies
mesh_terms: D021301:Antifreeze Proteins / Q000302:isolation & purification*; D007053:Ice; D012996:Solutions
chemical_list: D021301:Antifreeze Proteins; D007053:Ice; D012996:Solutions
keywords: Antifreeze protein; Ice affinity; Ice finger; Ice shell