Characterization of drug-protein interactions in blood using high-performance affinity chromatography

David S. Hage, Abby Jackson, Matthew R. Sobansky, John E. Schiel, Michelle J. Yoo, K. S. Joseph

Research output: Contribution to journalReview articlepeer-review

63 Scopus citations


The binding of drugs with proteins in blood, serum, or plasma is an important process in determining the activity, distribution, rate of excretion, and toxicity of drugs in the body. High-performance affinity chromatography (HPAC) has received a great deal of interest as a means for studying these interactions. This review examines the various techniques that have been used in HPAC to examine drug-protein binding and discusses the types of information that can be obtained through this approach. A comparison of these techniques with traditional methods for binding studies (e.g., equilibrium dialysis and ultrafiltration) will also be presented. The use of HPAC with specific serum proteins and binding agents will then be discussed, including HSA and α 1-acid glycoprotein (AGP). Several examples from the literature are provided to illustrate the applications of such research. Recent developments in this field are also described, such as the use of improved immobilization techniques, new data analysis methods, techniques for working directly with complex biological samples, and work with immobilized lipoproteins. The relative advantages and limitations of the methods that are described will be considered and the possible use of these techniques in the high-throughput screening or characterization of drug-protein binding will be discussed.

Original languageEnglish (US)
Pages (from-to)835-853
Number of pages19
JournalJournal of Separation Science
Issue number5-6
StatePublished - 2009


  • Drug-protein binding
  • High-performance affinity chromatography
  • Human serum albumin (HSA)
  • Lipoproteins
  • α-acid glycoprotein

ASJC Scopus subject areas

  • Analytical Chemistry
  • Filtration and Separation

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