Double coupling Edman chemistry for high-sensitivity automated protein sequencing.
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.
Ireland
I D
ID
Department of Chemistry, University of Alberta, Edmonton, Canada.
Lewis
D F
DF
Li
X F
XF
Renborg
A
A
Kwong
S
S
Chen
M
M
Dovichi
N J
NJ
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*
D000596:Amino Acids; D011506:Proteins; D010669:Phenylthiohydantoin; D016650:Fluorescein-5-isothiocyanate