Tags

Type your tag names separated by a space and hit enter

Capillary liquid chromatography of multiple peptides with on-line capillary electrophoresis immunoassay detection.
Electrophoresis. 2001 Oct; 22(17):3659-67.E

Abstract

A competitive immunoassay for neuropeptide Y (NPY) based on capillary electrophoresis (CE) with laser-induced fluorescence detection was developed utilizing polyclonal antisera as the immunoreagent and fluorescein-labeled NPY as the tracer. The assay was performed with on-line mixing of reagents, automated injections, and a 3 s separation time. The assay had a detection limit of 850 pM. To detect NPY at lower concentrations, the assay was coupled on-line to reversed-phase capillary liquid chromatography (LC). In this arrangement, 5 microL samples were preconcentrated by capillary LC and eluted by a gradient of isopropanol-containing mobile phase. The resulting chromatographic peaks were monitored by the CE immunoassay. With preconcentration, the concentration detection limit was improved to 40 microM and NPY could be measured in push-pull perfusion samples collected from the paraventricular nucleus of freely moving rats. The technique was extended to simultaneous detection of NPY and glucagon secretion from islets of Langerhans.

Authors+Show Affiliations

German I
Department of Chemistry, University of Florida, Gainesville, USA.
Roper MG
No affiliation info available
Kalra SP
No affiliation info available
Rhinehart E
No affiliation info available
Kennedy RT
No affiliation info available

MeSH

Chromatography, LiquidElectrophoresis, CapillaryImmunoassayNeuropeptide YOnline SystemsPeptidesSensitivity and Specificity

Pub Type(s)

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

Language

eng

PubMed ID

11699903

Citation

German, I, et al. "Capillary Liquid Chromatography of Multiple Peptides With On-line Capillary Electrophoresis Immunoassay Detection." Electrophoresis, vol. 22, no. 17, 2001, pp. 3659-67.
German I, Roper MG, Kalra SP, et al. Capillary liquid chromatography of multiple peptides with on-line capillary electrophoresis immunoassay detection. Electrophoresis. 2001;22(17):3659-67.
German, I., Roper, M. G., Kalra, S. P., Rhinehart, E., & Kennedy, R. T. (2001). Capillary liquid chromatography of multiple peptides with on-line capillary electrophoresis immunoassay detection. Electrophoresis, 22(17), 3659-67.
German I, et al. Capillary Liquid Chromatography of Multiple Peptides With On-line Capillary Electrophoresis Immunoassay Detection. Electrophoresis. 2001;22(17):3659-67. PubMed PMID: 11699903.
* Article titles in AMA citation format should be in sentence-case
TY - JOUR T1 - Capillary liquid chromatography of multiple peptides with on-line capillary electrophoresis immunoassay detection. AU - German,I, AU - Roper,M G, AU - Kalra,S P, AU - Rhinehart,E, AU - Kennedy,R T, PY - 2001/11/9/pubmed PY - 2002/4/3/medline PY - 2001/11/9/entrez SP - 3659 EP - 67 JF - Electrophoresis JO - Electrophoresis VL - 22 IS - 17 N2 - A competitive immunoassay for neuropeptide Y (NPY) based on capillary electrophoresis (CE) with laser-induced fluorescence detection was developed utilizing polyclonal antisera as the immunoreagent and fluorescein-labeled NPY as the tracer. The assay was performed with on-line mixing of reagents, automated injections, and a 3 s separation time. The assay had a detection limit of 850 pM. To detect NPY at lower concentrations, the assay was coupled on-line to reversed-phase capillary liquid chromatography (LC). In this arrangement, 5 microL samples were preconcentrated by capillary LC and eluted by a gradient of isopropanol-containing mobile phase. The resulting chromatographic peaks were monitored by the CE immunoassay. With preconcentration, the concentration detection limit was improved to 40 microM and NPY could be measured in push-pull perfusion samples collected from the paraventricular nucleus of freely moving rats. The technique was extended to simultaneous detection of NPY and glucagon secretion from islets of Langerhans. SN - 0173-0835 UR - https://www.unboundmedicine.com/medline/citation/11699903/Capillary_liquid_chromatography_of_multiple_peptides_with_on_line_capillary_electrophoresis_immunoassay_detection_ L2 - https://doi.org/10.1002/1522-2683(200109)22:17<3659::AID-ELPS3659>3.0.CO;2-I DB - PRIME DP - Unbound Medicine ER -
Grapherence [↓5]

Related Citations

More