Applications of capillary isoelectric focusing with liquid-core waveguide laser-induced fluorescence whole-column imaging detection.
Capillary isoelectric focusing (CIEF) with liquid-core waveguide (LCW) laser-induced fluorescence (LIF) whole-column imaging detection (WCID) is a recently developed high-resolution, high-sensitivity, and high-speed analytical tool for protein analysis. Several potential applications of this system were demonstrated in this study. First, this system was employed to separate naturally fluorescent phycobiliproteins. Second, denaturing CIEF was suggested to study the conformational and chemical microheterogeneity and to characterize proteins with identical pI values. Third, a modified noncovalent fluorescent labeling procedure was presented, which allows the simple and effective labeling of proteins, antibodies, and viruses with reduced multiple labeling and preserved activity. Finally, extracellular proteins were suggested as signaling biomarkers for evaluation of cell viability. The separation of cyanobacteria and their extracellular phycoerythrins was demonstrated. The effectiveness of CIEF-LCW-LIF-WCID for the analysis of proteins, antibodies, viruses, and cells has been illustrated.
Liu
Zhen
Z
Department of Chemistry, University of Waterloo, Waterloo, Ont., Canada N2L 3G1.
Pawliszyn
Janusz
J
D000906:Antibodies / Q000032:analysis; D002470:Cell Survival; D000458:Cyanobacteria / Q000737:chemistry; D005069:Evaluation Studies as Topic; D005453:Fluorescence; D005456:Fluorescent Dyes; D049452:Green Fluorescent Proteins / Q000032:analysis*; D007525:Isoelectric Focusing / Q000295:instrumentation / Q000379:methods*; D007834:Lasers; D045342:Light-Harvesting Protein Complexes / Q000302:isolation & purification; D008832:Microchemistry; D009930:Organic Chemicals; D011489:Protein Denaturation
D000906:Antibodies; D005456:Fluorescent Dyes; D045342:Light-Harvesting Protein Complexes; C433144:NanoOrange; D009930:Organic Chemicals; D049452:Green Fluorescent Proteins