title: Selective, bead-based global peptide capture using a bifunctional cross-linker.
abstract: Peptides are important species for a variety of biological functions. Detection and analysis of these molecules can be complicated by the presence of background matrix or contaminants. Therefore, a selective method to capture peptides could provide researchers with an option to isolate these remarkable species. Our goal was to perform a set of experiments that would validate the concept of a novel, selective peptide capture, whereby peptides are isolated on functionalized magnetic beads through the use of the heterobifunctional cross-linker, Sulfo-LC-SPDP. Matrix assisted laser desorption/ionization time-of-flight mass spectrometry (MALDI-TOF MS) was used to monitor mass changes associated with the cross-linking reaction. MALDI-TOF MS was then used to monitor conjugation between the cross-linked peptides and sulfhydryl magnetic beads by analyzing supernatant solutions for the presence or absence of cross-linked peptide. Through these experiments, we have proof of concept data confirming that peptides can be isolated on sulfhydryl magnetic beads by using Sulfo-LC-SPDP. This method is a suitable selective global peptide isolation strategy to separate the molecules from contaminating species or sample matrix. This novel method has a variety of potential applications and detection methods.
authors.affiliation: Department of Chemistry and Biochemistry, University of Notre Dame , 251 Nieuwland Science Hall, Notre Dame, Indiana 46556, United States.
authors.forename: Leigh A
authors.initials: LA
authors.lastname: Weston
authors.forename: Kerry M
authors.initials: KM
authors.lastname: Bauer
authors.forename: Susan B
authors.initials: SB
authors.lastname: Skube
authors.forename: Amanda B
authors.initials: AB
authors.lastname: Hummon
mesh_terms: D003110:Colonic Neoplasms / Q000378:metabolism*; D003432:Cross-Linking Reagents / Q000494:pharmacology*; D006801:Humans; D010446:Peptide Fragments / Q000032:analysis / Q000737:chemistry / Q000302:isolation & purification*; D019032:Spectrometry, Mass, Matrix-Assisted Laser Desorption-Ionization / Q000379:methods; D013388:Succinimides / Q000494:pharmacology*; D013438:Sulfhydryl Compounds / Q000737:chemistry*; D014407:Tumor Cells, Cultured
chemical_list: D003432:Cross-Linking Reagents; D010446:Peptide Fragments; D013388:Succinimides; D013438:Sulfhydryl Compounds; C018151:N-succinimidyl 3-(2-pyridyldithio)propionate