Purification and characterization of crystallins by aqueous two-phase extraction |
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Authors: | Ondrea Bermudez Daniel Forciniti |
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Institution: | (1) Chemical Engineering Department, University of Missouri-Rolla, 65409 Rolla, MO, USA |
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Abstract: | Crystallins are a family of water-soluble proteins that constitute up to 90% of the water-soluble proteins in mammalian eye
lenses. We present in this paper an alternative purification method for these proteins using polyethylene glycol/dextran aqueous
two-phase extraction. Under the appropriate conditions, we were able to recover the γ-crystallin fraction essentially free
of the remaining proteins. High concentrations of salt at a neutral pH maximize the recovery of γ-crystallins in the top phase
and minimize the contamination by the other proteins present in the lenses. The proposed protocol decreases the separation
time by about 50% The complex partition behavior observed for these proteins reflects a delicate balance between protein/phase-forming
species (various polymers and salts) and protein/protein interactions. This is evidenced in part, by the role played by the
largest proteins in this group as a “pseudo” phase-forming species. |
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Keywords: | crystallins purification aqueous two-phase systems |
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