Journal
BIOTECHNIQUES
Volume 68, Issue 3, Pages 148-154Publisher
FUTURE SCI LTD
DOI: 10.2144/btn-2019-0140
Keywords
automation; chromatography; dispersive solid-phase extraction; high-throughput screening; INtip chemistry; purification
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Automation gives researchers the ability to process and screen orders of magnitude higher numbers of samples than manual experimentation. Current biomacromolecule separation methodologies suffer from necessary manual intervention, making their translation to high-throughput automation difficult. Herein, we present the first characterization of biomacromolecule affinity purification via dispersive solid-phase extraction in a pipette tip (INtip). We use commercially available resin and compare efficiency with batch and spin column methodologies. Moreover, we measure the kinetics of binding and evaluate resin binding capacities. INtip technology is effective on, and scalable for, an automated platform (INTEGRA ASSIST). The results suggest that high-throughput biomolecular workflows will benefit from the integration of INtip separations. METHOD SUMMARY We have developed a pipette tip containing nickel-affinity resin that utilizes dispersive solid-phase extraction to purify histidine-tagged proteins at high yields and purity. This tip has been employed on an automated device and can be scaled for high-throughput applications. The efficiency, capacity and affinity of these tips were tested with three proteins in bacterial cell lysates.
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