Immobilization Of Alpha 1-Acid Glycoprotein For Chromatographic Studies Of Drug-Protein Binding Ii. Correction For Errors In Association Constant Measurements

Authors

Rangan Mallik, University of Nebraska-Lincoln
Hai Xuan, University of Nebraska-Lincoln
Georges Guiochon, University of Tennessee - KnoxvilleFollow
David S. Hage, University of Nebraska - LincolnFollow

Date of this Version

5-1-2008

Citation

Published in final edited form as: Anal Biochem. 2008 May 1; 376(1): 154–156. Version presented here is from NIH PubMed Central

Comments

Copyright Elsevier Inc. Used by permission

Abstract

A new method for the immobilization of α₁-acid glycoprotein (AGP) in HPLC columns was recently described for applications such as drug binding studies. Part of this earlier work used self-competition zonal elution studies to measure association equilibrium constants between immobilized AGP and R- or S-propranolol. It was later found that analysis of these data by a common equation derived for linear elution conditions gave erroneous values for experiments actually conducted under nonlinear conditions. This report discusses the nature of this error and uses frontal analysis to estimate the true binding strength between R- and S-propranolol and HPLC columns containing immobilized AGP.