The Effect of Guanidine Hydrochloride on Phase Diagram of PEG- Phosphate Aqueous Two-Phase System

This report focus on phase behavior of polyethylene glycol (PEG)4000/ phosphate/ guanidine hydrochloride/ water system at different guanidine hydrochloride concentrations and pH. The binodal of the systems was displaced toward higher concentrations of the components with increasing guanidine hydrochloride concentrations. The partition coefficient of guanidine hydrochloride was near unity and increased with decreasing pH and increasing PEG/salt (%w/w) ratio.

Authors:

• Farshad Rahimpour
• Mohsen Pirdashti

Keywords:

• Aqueous two-phase system
• guanidinehydrochloride
• partition coefficient
• phase diagram.

References:

[1] P. C. Singh and R. K. Singh, "Choosing an appropriate bioseparation
technique", Trend in Food science & Technology, vol.7, 1996.
[2] P. Å. Albartsson, Partition of cell particles and macromolecules, 3rd Ed.
New York: Willy, 1986.
[3] H. Walter, D. Brooks, and D. fisher, Partitioning in aqueous two-phase
systems, New York: Academic Press, 1985.
[4] F. Rahimpour., F. Feyzi., S. Maghsodi and R. H. Kaul, "Purification of
plasmid DNA with polymer-Salt aqueous two-phase system:
optimization using response surface methodology", Biotechnol. Bioeng.,
vol. 95, 2006, pp 627-637.
[5] F. Rahimpour., G. Mamo., F. Feyzi., S. Maghsoudi and R. H. Kaul,
"Optimization refolding and recovery of active recombinant bacillus
halodurans xylanase in polymer-salt aqueous two-phase system using
surface response analysis", J. Chromat. A, vol. 1141, 2007, pp. 32-40.
[6] E. D. B. Clark, "Protein refolding for industrial processes", Current
Opinion in Biotechnol., 2001, vol.12, pp. 202-207.
[7] R.A. Hart, P.M. Lester, D.H. Reifsnyder, J.R. Ogez, S.E. Builder,
"Large scale, In Situ isolation of periplasmic IGF-I from E. coli."
Bio./Technol. 12 (1994) 1113.
[8] D. Forciniti, "Protein refolding using aqueous two-phase systems" J.
Chromatogr. A 668 (1994) 95-99.
[9] C. Rämsch, L. B. Kleinelanghorst, E. A. Knieps, J. hommes and M.-R.
Kula, "Aqueous two-phase system containing urea; influence on phase
separation and stabilization of protein conformation by phase
components", Bitechnol. Prog., Vol. 15, 1999, pp. 493-499.
[10] C. Rämsch, L. B. Kleinelanghorst, E. A. Knieps., J. Thommes, and M. -
R. Kula, "Aqueous two-phase systems containing urea: influence of
protein Structure on Protein Partitioning", Biotechnol. Bioeng., vol. 69,
2000, pp. 83-90.
[11] P. Hagel., J. J. T. Gerding., W. Fieggen and H. Bloemendal, "Cyanate
formation in solutions of urea, I. Calculation of cyanate concentrations
at different temperature and pH", Biochim. Biophys. Acta, vol. 243,
1971, pp. 366-373.
[12] J. Cejka, Z. Vodražka and J. Salgk, "Carbamylation of globin in
electrophoresis and chromatography in the presence of urea", Biochim.
Biophys. Acta, vol. 154, 1968, pp. 589-591.
[13] A. Haghtalab and Mokhtarani, B.; "The new experimental data and a
new thermodynamic model based on group contribution for correlation
liquid- liquid equilibria in aqueous two-phase systems of PEG and
(K2HPO4 or Na2SO4)". Fluid Phase Equilibria, vol. 215, 2004, pp.
151-161.
[14] D. Estapé and U. Rinas, "Optimized procedures for purification and
solubilization of basic fibroblast growth factor inclusion bodies",
Biotechnol. Tech., vol.10, 1996, pp. 481-484.
[15] O. Annuziata., N. Asherie., A. Lomakin.,J. Pande., O. Ogun and G. B.
benedek, "Effect of polyethylene glycol on the liquid-liquid phase
transition in aqueous protein solutions" PNAS, vol.99, no.22, 2002, pp.
14165-14170.