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Stacking and separation of protein derivatives of naphthalene-2,3-dicarboxaldehyde by CE with light-emitting diode induced fluorescence detection.
Electrophoresis. 2008 Jan; 29(2):433-40.E

Abstract

We describe the stacking and separation of proteins by CE under discontinuous conditions in conjunction with light-emitting diode induced fluorescence (LEDIF) detection using a violet LED at 405 nm. The proteins were derivatized with naphthalene-2,3-dicarboxaldehyde (NDA) to form NDA-protein derivatives prior to CE-LEDIF analysis. During the separation, poly(ethylene oxide) (PEO) solution containing CTAB enters from the cathodic inlet to the capillary via electroosomotic flow (EOF). The optimum conditions are: the capillary was filled with 50 mM glycine buffer (pH 9.0) containing 1.0 mM CTAB, NDA-protein derivatives were prepared in deionized water containing 1.0 mM CTAB, and 0.6% PEO was prepared in 50 mM glycine (pH 9.0) containing 2.0 mM CTAB. The analysis of four NDA-protein derivatives is fast (<3 min), with RSD <1.5% in terms of migration time. In order to improve the sensitivity of NDA-protein derivatives, a stacking approach based on increases in viscosity and electric field, as well as sieving was applied. The efficient stacking approach provides LODs (S/N = 3) of 2.41, 0.59, 0.61, and 4.22 nM for trypsin inhibitor, HSA, beta-lactoglobulin, and lysozyme, respectively. In addition, we also applied the stacking approach to determination of the concentration of HSA in one urine sample, which was determined to be 0.31 +/- 0.05 microM (n = 3).

Authors+Show Affiliations

Chiu TC
Department of Chemistry, National Taiwan University, Taipei, Taiwan.
Tu WC
No affiliation info available
Chang HT
No affiliation info available

MeSH

AlbuminuriaCetrimoniumCetrimonium CompoundsElectroosmosisElectrophoresis, CapillaryFluorescenceHumansLactoglobulinsMuramidaseNaphthalenesOnline SystemsPolyethylene GlycolsProteinsSerum AlbuminTrypsin Inhibitors

Pub Type(s)

Journal Article
Research Support, Non-U.S. Gov't

Language

eng

PubMed ID

18064734

Citation

Chiu, Tai-Chia, et al. "Stacking and Separation of Protein Derivatives of Naphthalene-2,3-dicarboxaldehyde By CE With Light-emitting Diode Induced Fluorescence Detection." Electrophoresis, vol. 29, no. 2, 2008, pp. 433-40.
Chiu TC, Tu WC, Chang HT. Stacking and separation of protein derivatives of naphthalene-2,3-dicarboxaldehyde by CE with light-emitting diode induced fluorescence detection. Electrophoresis. 2008;29(2):433-40.
Chiu, T. C., Tu, W. C., & Chang, H. T. (2008). Stacking and separation of protein derivatives of naphthalene-2,3-dicarboxaldehyde by CE with light-emitting diode induced fluorescence detection. Electrophoresis, 29(2), 433-40.
Chiu TC, Tu WC, Chang HT. Stacking and Separation of Protein Derivatives of Naphthalene-2,3-dicarboxaldehyde By CE With Light-emitting Diode Induced Fluorescence Detection. Electrophoresis. 2008;29(2):433-40. PubMed PMID: 18064734.
* Article titles in AMA citation format should be in sentence-case
TY - JOUR T1 - Stacking and separation of protein derivatives of naphthalene-2,3-dicarboxaldehyde by CE with light-emitting diode induced fluorescence detection. AU - Chiu,Tai-Chia, AU - Tu,Wu-Chun, AU - Chang,Huan-Tsung, PY - 2007/12/8/pubmed PY - 2008/5/16/medline PY - 2007/12/8/entrez SP - 433 EP - 40 JF - Electrophoresis JO - Electrophoresis VL - 29 IS - 2 N2 - We describe the stacking and separation of proteins by CE under discontinuous conditions in conjunction with light-emitting diode induced fluorescence (LEDIF) detection using a violet LED at 405 nm. The proteins were derivatized with naphthalene-2,3-dicarboxaldehyde (NDA) to form NDA-protein derivatives prior to CE-LEDIF analysis. During the separation, poly(ethylene oxide) (PEO) solution containing CTAB enters from the cathodic inlet to the capillary via electroosomotic flow (EOF). The optimum conditions are: the capillary was filled with 50 mM glycine buffer (pH 9.0) containing 1.0 mM CTAB, NDA-protein derivatives were prepared in deionized water containing 1.0 mM CTAB, and 0.6% PEO was prepared in 50 mM glycine (pH 9.0) containing 2.0 mM CTAB. The analysis of four NDA-protein derivatives is fast (<3 min), with RSD <1.5% in terms of migration time. In order to improve the sensitivity of NDA-protein derivatives, a stacking approach based on increases in viscosity and electric field, as well as sieving was applied. The efficient stacking approach provides LODs (S/N = 3) of 2.41, 0.59, 0.61, and 4.22 nM for trypsin inhibitor, HSA, beta-lactoglobulin, and lysozyme, respectively. In addition, we also applied the stacking approach to determination of the concentration of HSA in one urine sample, which was determined to be 0.31 +/- 0.05 microM (n = 3). SN - 0173-0835 UR - https://www.unboundmedicine.com/medline/citation/18064734/Stacking_and_separation_of_protein_derivatives_of_naphthalene_23_dicarboxaldehyde_by_CE_with_light_emitting_diode_induced_fluorescence_detection_ DB - PRIME DP - Unbound Medicine ER -