Affinity surfactants as reversibly bound ligands for high-performance affinity chromatography

J. L. Torres, Roberto Z Guzman, R. G. Carbonell, P. K. Kilpatrick

Research output: Contribution to journalArticle

15 Citations (Scopus)

Abstract

Pyridine was coupled covalently to a nonionic ethoxylated alcohol: octaethylene glycol n-hexadecyl ether. This modified surfactant was found to be a reversible, competitive inhibitor of horse serum cholinesterase. The surfactant bound irreversibly, in aqueous media, to octadecylbonded reverse phase silica particles commonly used for high-performance liquid chromatography. The amount of ligand bound was found to be 550 μmol/ml of packing, a concentration that is over 100 times higher than what can be normally bound to agarose affinity chromatography supports. With this packing, a 280-fold purification of cholinesterase from horse serum and a 79-fold purification of human serum cholinesterase were accomplished, with yields greater than 80%, using a 2-cm-long column and a 7-min elution time. The affinity surfactant could be eluted from the column using a 6:4 (v/v) mixture of methanol and isopropanol. This technique should be generally applicable in the development of biospecific supports for high-performance affinity chromatography.

Original languageEnglish (US)
Pages (from-to)411-418
Number of pages8
JournalAnalytical Biochemistry
Volume171
Issue number2
DOIs
StatePublished - 1988
Externally publishedYes

Fingerprint

Affinity chromatography
Cholinesterases
Affinity Chromatography
Surface-Active Agents
Ligands
Horses
Purification
Serum
Agarose Chromatography
Glycols
2-Propanol
High performance liquid chromatography
Silicon Dioxide
Ether
Sepharose
Methanol
High Pressure Liquid Chromatography
Alcohols

Keywords

  • affinity-labeled surfactants
  • high-performance affinity chromatography
  • ligand immobilization
  • Nonionic surfactants
  • octadecyl-bonded silica
  • serum cholinesterase

ASJC Scopus subject areas

  • Biochemistry
  • Biophysics
  • Molecular Biology

Cite this

Affinity surfactants as reversibly bound ligands for high-performance affinity chromatography. / Torres, J. L.; Guzman, Roberto Z; Carbonell, R. G.; Kilpatrick, P. K.

In: Analytical Biochemistry, Vol. 171, No. 2, 1988, p. 411-418.

Research output: Contribution to journalArticle

Torres, J. L. ; Guzman, Roberto Z ; Carbonell, R. G. ; Kilpatrick, P. K. / Affinity surfactants as reversibly bound ligands for high-performance affinity chromatography. In: Analytical Biochemistry. 1988 ; Vol. 171, No. 2. pp. 411-418.
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