Format

Send to

Choose Destination
Cryobiology. 2016 Jun;72(3):258-63. doi: 10.1016/j.cryobiol.2016.03.009. Epub 2016 Mar 26.

Ice-shell purification of ice-binding proteins.

Author information

1
Department of Biochemistry, and Genetics Otago, University of Otago, PO Box 56, Dunedin, 9054, New Zealand. Electronic address: craig.marshall@otago.ac.nz.
2
Department of Biomedical and Molecular Sciences, Queen's University, Kingston, Ontario, K7L 3N6, Canada.

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.

KEYWORDS:

Antifreeze protein; Ice affinity; Ice finger; Ice shell

PMID:
27025155
DOI:
10.1016/j.cryobiol.2016.03.009
[Indexed for MEDLINE]

Supplemental Content

Full text links

Icon for Elsevier Science
Loading ...
Support Center