Solid-liquid equilibrium for proteins in solutions with an unconventional salt (ammonium carbamate): Phase behavior analysis

Solid-liquid equilibrium for proteins in solutions with an unconventional salt (ammonium carbamate): Phase behavior analysis

Author Medeiros Hirata, Gisele Atsuko Autor UNIFESP Google Scholar
Pessoa Filho, Pedro de Alcantara Google Scholar
Miranda, Everson Alves Google Scholar
Abstract In this manuscript, we report the precipitation and crystallization of three proteins (chicken egg-white lysozyme, porcine insulin and bovine insulin) using ammonium carbamate as salting-out agent. For these proteins, data on solubility, metastability limits, and osmotic second virial coefficient (B-22) as a function of salt concentration at constant temperature (25.0 degrees C) were obtained. The value of osmotic second virial coefficient (B-22) can be regarded as a selection criterion for protein crystallization, as it is related to the interaction potential between protein molecules. Negative values of B-22 and large values of the salting out constant showed that ammonium carbamate is a good precipitating agent for these proteins. Crystallization trials conducted under specific conditions showed that both insulins form amorphous precipitates in ammonium carbamate solutions, which is compatible with the large negative values of B-22 measured. Conversely, lysozyme crystals were obtained under all conditions studied, and B-22 values for this enzyme were within or close to the crystallization slot. (C) 2017 Elsevier B.V. All rights reserved.
Keywords Protein
Crystallization
Osmotic second virial coefficient
Volatile salt
xmlui.dri2xhtml.METS-1.0.item-coverage Amsterdam
Language English
Sponsor Brazilian agency CNPq
Brazilian agency CAPES
Brazilian agency FAEPEX-UNI-CAMP
Brazilian agency FAPESP
Grant number CNPq
CAPES
FAEPEX-UNI-CAMP
FAPESP: 2010/52524-5
Date 2017
Published in Fluid Phase Equilibria. Amsterdam, v. 443, p. 1-8, 2017.
ISSN 0378-3812 (Sherpa/Romeo, impact factor)
Publisher Elsevier Science Bv
Extent 1-8
Origin http://dx.doi.org/10.1016/j.fluid.2017.03.031
Access rights ACESSO RESTRITO
Type Article
Web of Science ID WOS:000401688500002
URI https://repositorio.unifesp.br/handle/11600/53448

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