Automated affinity capture-release of biotin-containing conjugates using a lab-on-valve apparatus coupled to UV/visible and electrospray ionization mass spectrometry.
We report a new method for automated affinity capture and release of biotin-containing conjugates on immobilized streptavidin using a lab-on-valve (LOV) bead injection apparatus. The apparatus is also coupled to UV/visible and electrospray ionization mass spectrometry (ESI-MS) for monitoring the captured and released biotin-containing conjugates. Dissociation rate constants for release from streptavidin of two chromophore-tagged biotin conjugates were measured by UV/visible spectrometry and the dissociation was simultaneously monitored by ESI-MS. The LOV-ESI-MS instrument was also used for repetitive assays of lysosomal beta-galactosidase in human cell homogenates. Fast analysis in 4.5 min/full cycle and robust operation in 60 repetitive analyses are demonstrated that are promising for transfer of the LOV-ESI-MS technology into clinical practice.
D000345:Affinity Labels; D001710:Biotin / Q000737:chemistry*; D009682:Magnetic Resonance Spectroscopy; D021241:Spectrometry, Mass, Electrospray Ionization / Q000295:instrumentation*; D013056:Spectrophotometry, Ultraviolet / Q000295:instrumentation*; D001616:beta-Galactosidase / Q000378:metabolism
D000345:Affinity Labels; D001710:Biotin; D001616:beta-Galactosidase