1H high-resolution magic-angle spinning (HR-MAS) NMR analysis of ligand density on resins using a resin internal standard.
We recently attempted to generate an affinity chromatography adsorbent to purify cytochrome P450 4A1 by coupling 11-(1'-imidazolyl)-3,6,9-trioxaundecanoic acid to Toyopearl AF-Amino 650 M resin. Variations in ligand density for several resin batches were quantified by high-resolution magic-angle spinning (HR-MAS) NMR spectroscopy using a novel resin internal standard. The uniquely designed ImQ internal resin standard yields its signature resonance in a transparent region of the analyte spectrum making suppression of the polymer background unnecessary. This method enabled us to target a reasonable ligand density for enzyme purification and provides an advantageous alternative to quantitation against soluble standards or protonated solvent.
Lucas
Laura H
LH
Department of Chemistry, University of Kansas, 1251 Wescoe Hall Drive, 2010 Malott Hall, Lawrence, KS 66045-7582, USA.
Cerny
Matthew A
MA
Koen
Yakov M
YM
Hanzlik
Robert P
RP
Larive
Cynthia K
CK
D000818:Animals; D002846:Chromatography, Affinity / Q000295:instrumentation* / Q000379:methods; D008024:Ligands; D009682:Magnetic Resonance Spectroscopy / Q000379:methods*; D008745:Methylation; D015394:Molecular Structure; D051381:Rats; D012015:Reference Standards; D012117:Resins, Synthetic / Q000032:analysis* / Q000737:chemistry*; D014316:Tritium / Q000737:chemistry
D008024:Ligands; D012117:Resins, Synthetic; D014316:Tritium