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Article: Feasibility study of enzyme-amplified sandwich immunoassay using protein G capillary affinity chromatography and laser induced fluorescence detection

TitleFeasibility study of enzyme-amplified sandwich immunoassay using protein G capillary affinity chromatography and laser induced fluorescence detection
Authors
Issue Date2001
PublisherTaylor & Francis Inc. The Journal's web site is located at http://www.tandf.co.uk/journals/titles/10826076.asp
Citation
Journal Of Liquid Chromatography And Related Technologies, 2001, v. 24 n. 13, p. 1953-1963 How to Cite?
AbstractThe feasibility of performing the enzyme-amplified sandwich immunoassay with protein G capillary affinity chromatography and laser induced fluorescence (LIF) detection was investigated using the determination of human immunoglobulin G (hIgG) as a model system. The incubated mixture of samples containing hIgG and alkaline phosphatase (ALP) conjugated goat anti-human IgG F(ab')2 fragment was loaded onto the capillary column packed with recombinant protein G bound perfusive support in neutral pH. After nonretained compounds were eluted, the fluorogenic ALP substrate, fluorescein diphosphate (FDP), was loaded onto the capillary column followed by stop-flow incubation. Finally, the product of ALP-mediated hydrolysis of FDP, fluorescein, was swept out of the capillary column and detected with a LIF detector using the 488 nm line of an argon ion laser as the excitation source. Chromatographic conditions were optimized. The calibration curve for hIgG was linear over the range of 0.5-50 pmol/L (r2=0.999) with the limit of detection of 0.2 pmol/L.
Persistent Identifierhttp://hdl.handle.net/10722/173251
ISSN
2015 Impact Factor: 0.669
2015 SCImago Journal Rankings: 0.299
ISI Accession Number ID
References

 

DC FieldValueLanguage
dc.contributor.authorWang, Qen_US
dc.contributor.authorWang, Yen_US
dc.contributor.authorLuo, Gen_US
dc.contributor.authorYeung, WSBen_US
dc.date.accessioned2012-10-30T06:28:48Z-
dc.date.available2012-10-30T06:28:48Z-
dc.date.issued2001en_US
dc.identifier.citationJournal Of Liquid Chromatography And Related Technologies, 2001, v. 24 n. 13, p. 1953-1963en_US
dc.identifier.issn1082-6076en_US
dc.identifier.urihttp://hdl.handle.net/10722/173251-
dc.description.abstractThe feasibility of performing the enzyme-amplified sandwich immunoassay with protein G capillary affinity chromatography and laser induced fluorescence (LIF) detection was investigated using the determination of human immunoglobulin G (hIgG) as a model system. The incubated mixture of samples containing hIgG and alkaline phosphatase (ALP) conjugated goat anti-human IgG F(ab')2 fragment was loaded onto the capillary column packed with recombinant protein G bound perfusive support in neutral pH. After nonretained compounds were eluted, the fluorogenic ALP substrate, fluorescein diphosphate (FDP), was loaded onto the capillary column followed by stop-flow incubation. Finally, the product of ALP-mediated hydrolysis of FDP, fluorescein, was swept out of the capillary column and detected with a LIF detector using the 488 nm line of an argon ion laser as the excitation source. Chromatographic conditions were optimized. The calibration curve for hIgG was linear over the range of 0.5-50 pmol/L (r2=0.999) with the limit of detection of 0.2 pmol/L.en_US
dc.languageengen_US
dc.publisherTaylor & Francis Inc. The Journal's web site is located at http://www.tandf.co.uk/journals/titles/10826076.aspen_US
dc.relation.ispartofJournal of Liquid Chromatography and Related Technologiesen_US
dc.titleFeasibility study of enzyme-amplified sandwich immunoassay using protein G capillary affinity chromatography and laser induced fluorescence detectionen_US
dc.typeArticleen_US
dc.identifier.emailYeung, WSB:wsbyeung@hkucc.hku.hken_US
dc.identifier.authorityYeung, WSB=rp00331en_US
dc.description.naturelink_to_subscribed_fulltexten_US
dc.identifier.doi10.1081/JLC-100104437en_US
dc.identifier.scopuseid_2-s2.0-0034909949en_US
dc.relation.referenceshttp://www.scopus.com/mlt/select.url?eid=2-s2.0-0034909949&selection=ref&src=s&origin=recordpageen_US
dc.identifier.volume24en_US
dc.identifier.issue13en_US
dc.identifier.spage1953en_US
dc.identifier.epage1963en_US
dc.identifier.isiWOS:000170103300007-
dc.publisher.placeUnited Statesen_US
dc.identifier.scopusauthoridWang, Q=7406915351en_US
dc.identifier.scopusauthoridWang, Y=13310049900en_US
dc.identifier.scopusauthoridLuo, G=7401536462en_US
dc.identifier.scopusauthoridYeung, WSB=7102370745en_US

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