Analysis of drug-protein binding by ultrafast affinity chromatography using immobilized human serum albumin

Rangan Mallik, Michelle J. Yoo, Chad J. Briscoe, David S. Hage

Research output: Contribution to journalArticle

53 Citations (Scopus)

Abstract

Human serum albumin (HSA) was explored for use as a stationary phase and ligand in affinity microcolumns for the ultrafast extraction of free drug fractions and the use of this information for the analysis of drug-protein binding. Warfarin, imipramine, and ibuprofen were used as model analytes in this study. It was found that greater than 95% extraction of all these drugs could be achieved in as little as 250ms on HSA microcolumns. The retained drug fraction was then eluted from the same column under isocratic conditions, giving elution in less than 40 s when working at 4.5 mL/min. The chromatographic behavior of this system gave a good fit with that predicted by computer simulations based on a reversible, saturable model for the binding of an injected drug with immobilized HSA. The free fractions measured by this method were found to be comparable to those determined by ultrafiltration, and equilibrium constants estimated by this approach gave good agreement with literature values. Advantages of this method include its speed and the relatively low cost of microcolumns that contain HSA. The ability of HSA to bind many types of drugs also creates the possibility of using the same affinity microcolumn to study and measure the free fractions for a variety of pharmaceutical agents. These properties make this technique appealing for use in drug-binding studies and in the high-throughput screening of new drug candidates.

Original languageEnglish (US)
Pages (from-to)2796-2803
Number of pages8
JournalJournal of Chromatography A
Volume1217
Issue number17
DOIs
StatePublished - Apr 2010

Fingerprint

Affinity chromatography
Affinity Chromatography
Protein Binding
Serum Albumin
Pharmaceutical Preparations
Preclinical Drug Evaluations
Imipramine
Ibuprofen
Ultrafiltration
Warfarin
Equilibrium constants
Computer Simulation
Screening
Ligands
Throughput
Costs and Cost Analysis

Keywords

  • Affinity chromatography
  • Affinity microcolumns
  • Drug-protein binding
  • High-throughput screening
  • Human serum albumin
  • Ibuprofen
  • Imipramine
  • Warfarin

ASJC Scopus subject areas

  • Analytical Chemistry
  • Biochemistry
  • Organic Chemistry

Cite this

Analysis of drug-protein binding by ultrafast affinity chromatography using immobilized human serum albumin. / Mallik, Rangan; Yoo, Michelle J.; Briscoe, Chad J.; Hage, David S.

In: Journal of Chromatography A, Vol. 1217, No. 17, 04.2010, p. 2796-2803.

Research output: Contribution to journalArticle

Mallik, Rangan ; Yoo, Michelle J. ; Briscoe, Chad J. ; Hage, David S. / Analysis of drug-protein binding by ultrafast affinity chromatography using immobilized human serum albumin. In: Journal of Chromatography A. 2010 ; Vol. 1217, No. 17. pp. 2796-2803.
@article{d6b32fd4c5094703b6f3047da5426996,
title = "Analysis of drug-protein binding by ultrafast affinity chromatography using immobilized human serum albumin",
abstract = "Human serum albumin (HSA) was explored for use as a stationary phase and ligand in affinity microcolumns for the ultrafast extraction of free drug fractions and the use of this information for the analysis of drug-protein binding. Warfarin, imipramine, and ibuprofen were used as model analytes in this study. It was found that greater than 95{\%} extraction of all these drugs could be achieved in as little as 250ms on HSA microcolumns. The retained drug fraction was then eluted from the same column under isocratic conditions, giving elution in less than 40 s when working at 4.5 mL/min. The chromatographic behavior of this system gave a good fit with that predicted by computer simulations based on a reversible, saturable model for the binding of an injected drug with immobilized HSA. The free fractions measured by this method were found to be comparable to those determined by ultrafiltration, and equilibrium constants estimated by this approach gave good agreement with literature values. Advantages of this method include its speed and the relatively low cost of microcolumns that contain HSA. The ability of HSA to bind many types of drugs also creates the possibility of using the same affinity microcolumn to study and measure the free fractions for a variety of pharmaceutical agents. These properties make this technique appealing for use in drug-binding studies and in the high-throughput screening of new drug candidates.",
keywords = "Affinity chromatography, Affinity microcolumns, Drug-protein binding, High-throughput screening, Human serum albumin, Ibuprofen, Imipramine, Warfarin",
author = "Rangan Mallik and Yoo, {Michelle J.} and Briscoe, {Chad J.} and Hage, {David S.}",
year = "2010",
month = "4",
doi = "10.1016/j.chroma.2010.02.026",
language = "English (US)",
volume = "1217",
pages = "2796--2803",
journal = "Journal of Chromatography A",
issn = "0021-9673",
number = "17",

}

TY - JOUR

T1 - Analysis of drug-protein binding by ultrafast affinity chromatography using immobilized human serum albumin

AU - Mallik, Rangan

AU - Yoo, Michelle J.

AU - Briscoe, Chad J.

AU - Hage, David S.

PY - 2010/4

Y1 - 2010/4

N2 - Human serum albumin (HSA) was explored for use as a stationary phase and ligand in affinity microcolumns for the ultrafast extraction of free drug fractions and the use of this information for the analysis of drug-protein binding. Warfarin, imipramine, and ibuprofen were used as model analytes in this study. It was found that greater than 95% extraction of all these drugs could be achieved in as little as 250ms on HSA microcolumns. The retained drug fraction was then eluted from the same column under isocratic conditions, giving elution in less than 40 s when working at 4.5 mL/min. The chromatographic behavior of this system gave a good fit with that predicted by computer simulations based on a reversible, saturable model for the binding of an injected drug with immobilized HSA. The free fractions measured by this method were found to be comparable to those determined by ultrafiltration, and equilibrium constants estimated by this approach gave good agreement with literature values. Advantages of this method include its speed and the relatively low cost of microcolumns that contain HSA. The ability of HSA to bind many types of drugs also creates the possibility of using the same affinity microcolumn to study and measure the free fractions for a variety of pharmaceutical agents. These properties make this technique appealing for use in drug-binding studies and in the high-throughput screening of new drug candidates.

AB - Human serum albumin (HSA) was explored for use as a stationary phase and ligand in affinity microcolumns for the ultrafast extraction of free drug fractions and the use of this information for the analysis of drug-protein binding. Warfarin, imipramine, and ibuprofen were used as model analytes in this study. It was found that greater than 95% extraction of all these drugs could be achieved in as little as 250ms on HSA microcolumns. The retained drug fraction was then eluted from the same column under isocratic conditions, giving elution in less than 40 s when working at 4.5 mL/min. The chromatographic behavior of this system gave a good fit with that predicted by computer simulations based on a reversible, saturable model for the binding of an injected drug with immobilized HSA. The free fractions measured by this method were found to be comparable to those determined by ultrafiltration, and equilibrium constants estimated by this approach gave good agreement with literature values. Advantages of this method include its speed and the relatively low cost of microcolumns that contain HSA. The ability of HSA to bind many types of drugs also creates the possibility of using the same affinity microcolumn to study and measure the free fractions for a variety of pharmaceutical agents. These properties make this technique appealing for use in drug-binding studies and in the high-throughput screening of new drug candidates.

KW - Affinity chromatography

KW - Affinity microcolumns

KW - Drug-protein binding

KW - High-throughput screening

KW - Human serum albumin

KW - Ibuprofen

KW - Imipramine

KW - Warfarin

UR - http://www.scopus.com/inward/record.url?scp=77953693949&partnerID=8YFLogxK

UR - http://www.scopus.com/inward/citedby.url?scp=77953693949&partnerID=8YFLogxK

U2 - 10.1016/j.chroma.2010.02.026

DO - 10.1016/j.chroma.2010.02.026

M3 - Article

C2 - 20227701

AN - SCOPUS:77953693949

VL - 1217

SP - 2796

EP - 2803

JO - Journal of Chromatography A

JF - Journal of Chromatography A

SN - 0021-9673

IS - 17

ER -