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Electroimmobilization of proinsulin C-peptide to a quartz crystal microbalance sensor chip for protein affinity purification
Authors:Melles Ermias  Anderson Henrik  Wallinder Daniel  Shafqat Jawed  Bergman Tomas  Aastrup Teodor  Jörnvall Hans
Institution:Department of Medical Biochemistry and Biophysics, Karolinska Institutet, SE-171 77 Stockholm, Sweden.
Abstract:Proinsulin C-peptide was electroimmobilized to a quartz crystal microbalance sensor chip, localizing this low-pI peptide for covalent attachment to activated surface carboxyl groups. The resulting chip was used in a continuous flow biosensor to capture anti-C-peptide antibodies, which could subsequently be eluted in 5% formic acid between air bubbles for efficient recovery and mass spectrometric identification. The method is reproducible through repeated cycles, providing affinity purification of proteins under real-time monitoring of the binding and elution processes.
Keywords:Electroimmobilization  Quartz crystal microbalance chip  Biosensor  Proinsulin C-peptide  Mass spectrometry
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