title: Double coupling Edman chemistry for high-sensitivity automated protein sequencing.
abstract: Capillary electrophoresis with laser-induced fluorescence detection (CE-LIF) is a promising new method for the analysis of protein sequencing products. It gives 10 zmol (1 zmol = 10(-21) mol) limits of detection (3 sigma) for fluorescein thiohydantoin (FTH) amino acids. We have developed a separation for the (FTH)-amino acid products generated from 18 of the 20 coded amino acids. The extremely low volume requirement associated with CE-LIF makes it incompatible with commercial sequencers. For this reason, we have also been developing a miniaturized sequencer that can be more easily coupled to our detection system. Both the CE-LIF system and the miniaturized sequencer are described.
authors.affiliation: Department of Chemistry, University of Alberta, Edmonton, Canada.
authors.forename: I D
authors.initials: ID
authors.lastname: Ireland
authors.forename: D F
authors.initials: DF
authors.lastname: Lewis
authors.forename: X F
authors.initials: XF
authors.lastname: Li
authors.forename: A
authors.initials: A
authors.lastname: Renborg
authors.forename: S
authors.initials: S
authors.lastname: Kwong
authors.forename: M
authors.initials: M
authors.lastname: Chen
authors.forename: N J
authors.initials: NJ
authors.lastname: Dovichi
mesh_terms: D000595:Amino Acid Sequence; D000596:Amino Acids / Q000032:analysis; D016650:Fluorescein-5-isothiocyanate / Q000737:chemistry*; D005453:Fluorescence; D010669:Phenylthiohydantoin / Q000737:chemistry*; D011506:Proteins / Q000032:analysis* / Q000737:chemistry; D012680:Sensitivity and Specificity; D017421:Sequence Analysis / Q000379:methods*
chemical_list: D000596:Amino Acids; D011506:Proteins; D010669:Phenylthiohydantoin; D016650:Fluorescein-5-isothiocyanate