Pulsed ultrafiltration mass spectrometry: a new method for screening combinatorial libraries

Anal Chem. 1997 Jun 1;69(11):2159-64. doi: 10.1021/ac970132j.

Abstract

In response to the need for rapid screening of combinatorial libraries to identify new lead compounds during drug discovery, we have developed an on-line combination of ultrafiltration and electrospray mass spectrometry, called pulsed ultrafiltration mass spectrometry, which facilitates the identification of solution-phase ligands in library mixtures that bind to solution-phase receptors. After ligands contained in a library mixture were bound to a macromolecular receptor, e.g., human serum albumin or calf intestine adenosine deaminase, the ligand-receptor complexes were purified by ultrafiltration and then dissociated using methanol to elute the ligands into the electrospray mass spectrometer for detection. Ligands with dissociation constants in the micromolar to nanomolar range were successfully bound, released, and detected using this method, including warfarin, salicylate, furosemide, and thyroxine binding to human serum albumin, and erythro-9-(2-hydroxy-3-nonyl)adenine binding to calf intestine adenosine deaminase. Repetitive bind- and-release experiments demonstrated that the receptor could be reused. Thus, pulsed ultrafiltration mass spectrometry was shown to provide a simple and powerful new method for the screening of combinatorial libraries in support of new drug discovery.

MeSH terms

  • Adenine / analogs & derivatives
  • Adenine / analysis
  • Adenine / metabolism
  • Adenosine Deaminase / metabolism*
  • Animals
  • Cattle
  • Drug Design
  • Enzyme Inhibitors / analysis
  • Enzyme Inhibitors / metabolism
  • Furosemide / analysis
  • Furosemide / metabolism
  • Humans
  • Intestines / enzymology
  • Ligands
  • Mass Spectrometry / methods*
  • Methanol / chemistry
  • Molecular Weight
  • Online Systems
  • Pulsatile Flow
  • Salicylates / analysis
  • Salicylates / metabolism
  • Salicylic Acid
  • Serum Albumin / metabolism*
  • Thyroxine / metabolism
  • Ultrafiltration
  • Warfarin / analysis
  • Warfarin / metabolism

Substances

  • Enzyme Inhibitors
  • Ligands
  • Salicylates
  • Serum Albumin
  • 9-(2-hydroxy-3-nonyl)adenine
  • Warfarin
  • Furosemide
  • Adenosine Deaminase
  • Adenine
  • Salicylic Acid
  • Thyroxine
  • Methanol