Electroimmobilization of proinsulin C-peptide to a quartz crystal microbalance sensor chip for protein affinity purification.
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.
Melles
Ermias
E
Department of Medical Biochemistry and Biophysics, Karolinska Institutet, SE-171 77 Stockholm, Sweden.
Anderson
Henrik
H
Wallinder
Daniel
D
Shafqat
Jawed
J
Bergman
Tomas
T
Aastrup
Teodor
T
Jörnvall
Hans
H
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
D002096:C-Peptide; D011791:Quartz; D011384:Proinsulin