title: Electroimmobilization of proinsulin C-peptide to a quartz crystal microbalance sensor chip for protein affinity purification.
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.
authors.affiliation: Department of Medical Biochemistry and Biophysics, Karolinska Institutet, SE-171 77 Stockholm, Sweden.
authors.forename: Ermias
authors.initials: E
authors.lastname: Melles
authors.forename: Henrik
authors.initials: H
authors.lastname: Anderson
authors.forename: Daniel
authors.initials: D
authors.lastname: Wallinder
authors.forename: Jawed
authors.initials: J
authors.lastname: Shafqat
authors.forename: Tomas
authors.initials: T
authors.lastname: Bergman
authors.forename: Teodor
authors.initials: T
authors.lastname: Aastrup
authors.forename: Hans
authors.initials: H
authors.lastname: Jörnvall
mesh_terms: D000595:Amino Acid Sequence; D001666:Binding Sites, Antibody; D015374:Biosensing Techniques / Q000295:instrumentation*; D002096:C-Peptide / Q000737:chemistry / Q000302:isolation & purification*; D006801:Humans; D008969:Molecular Sequence Data; D011384:Proinsulin / Q000737:chemistry / Q000302:isolation & purification*; D011485:Protein Binding; D011791:Quartz; D019032:Spectrometry, Mass, Matrix-Assisted Laser Desorption-Ionization
chemical_list: D002096:C-Peptide; D011791:Quartz; D011384:Proinsulin