TY - JOUR
T1 - Anion-exchange chromatographic behavior of recombinant rat cytochrome b5. Thermodynamic driving forces and temperature dependence of the stoichiometric displacement parameter Z
AU - Roush, David J.
AU - Gill, Davinder S.
AU - Willson, Richard C.
N1 - Funding Information:
E. coli strain TBl expressing recombinant soluble core of rat cytochromeb , was the generous gift of Dr. Stephen Sligar of the University of Illinois. We would like to thank Dr. Sligar and Dr. Karla Rodgers for helpful discussions.S up-port was provided by NSF under CTS-8910087, and by an NSF Presidential Young Investigator Award to R.C.W. Additional support was provided by Pharmaciaa nd by the Waters Chroma-tographyD ivision of Millipore Corporation.
Copyright:
Copyright 2014 Elsevier B.V., All rights reserved.
PY - 1993/11/5
Y1 - 1993/11/5
N2 - The HPLC anion-exchange isocratic retention behavior of the recombinant soluble core of wild type rat cytochrome b5 on Mono Q HR 5/5 was investigated as a function of temperature and sodium chloride concentration at fixed eluent flow-rates. Retention was measured over a range of eluent flow-rates at a specified temperature to determine if true adsorption equilibrium could be approximated by the HPLC method. Apparent Van't Hoff enthalpies of adsorption obtained from the HPLC retention data were positive, indicating an entropically driven spontaneous adsorption process, and were found to decline with increasing ionic strength. The retention results were interpreted in terms of the stoichiometric displacement model to obtain the apparent number of binding sites in the contact region, Z, as a function of temperature and of protein concentration. Z was found to depend significantly on temperature, even under conditions of nearly complete protein recovery, but did not depend on protein concentration at the low loadings studied.
AB - The HPLC anion-exchange isocratic retention behavior of the recombinant soluble core of wild type rat cytochrome b5 on Mono Q HR 5/5 was investigated as a function of temperature and sodium chloride concentration at fixed eluent flow-rates. Retention was measured over a range of eluent flow-rates at a specified temperature to determine if true adsorption equilibrium could be approximated by the HPLC method. Apparent Van't Hoff enthalpies of adsorption obtained from the HPLC retention data were positive, indicating an entropically driven spontaneous adsorption process, and were found to decline with increasing ionic strength. The retention results were interpreted in terms of the stoichiometric displacement model to obtain the apparent number of binding sites in the contact region, Z, as a function of temperature and of protein concentration. Z was found to depend significantly on temperature, even under conditions of nearly complete protein recovery, but did not depend on protein concentration at the low loadings studied.
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U2 - 10.1016/0021-9673(93)83176-S
DO - 10.1016/0021-9673(93)83176-S
M3 - Article
C2 - 8269055
AN - SCOPUS:0027331420
VL - 653
SP - 207
EP - 218
JO - Journal of Chromatography A
JF - Journal of Chromatography A
SN - 0021-9673
IS - 2
ER -