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Article: Multi-dimension microchip-capillary electrophoresis device for determination of functional proteins in infant milk formula.

TitleMulti-dimension microchip-capillary electrophoresis device for determination of functional proteins in infant milk formula.
Authors
Issue Date2013
PublisherElsevier BV. The Journal's web site is located at http://www.elsevier.com/locate/chroma
Citation
Journal of Chromatography A, 2013, v. 1304, p. 220-226 How to Cite?
AbstractTo improve resolution of important minor proteins and eliminate time-consuming precipitation of major protein with associated analyte co-precipitation risk, a multi-dimension strategy is adopted in the 2D microchip-CE device to isolate major proteins on-chip, enrich minor proteins in capillary before their separation in CE for UV quantitation. A standard fluorescent protein mixture containing FITC-BSA, myoglobin and cytochrome as specific pI markers has prepared to demonstrate capability of the device to fractionate minor proteins by IEF. The results using a standard protein mixture with profile resembling infant milk formula show a complete isolation of high abundance proteins by a 2-min 1D IEF run. The subsequent t-ITP/CZE run by on-chip high voltage switching delivers a high stacking ratio, realizing 60 folds enrichment of isolated protein fractions. All five important functional proteins (LF, IgG, alpha-LA, beta-LgA and beta-LgB) known to fortify infant milk formula are isolated and determined using two consecutive t-ITP-CZE runs within a 18-min total assay time, a significant saving compared to several hours conventional pretreatment. For a 100g infant milk formula sample, working ranges of 20-8000mg, repeatability 3.8-5.3% and detection limits 2.3-10mg have been achieved to meet government regulations. Method reliability is established by 100% recoveries and agreeable results within expected ranges and labeled values. The capability of the device for field operation, rapid assay with quick results, label-free universal detection, simple operation by aqueous dissolution before injection, and the demanding matching in 2D separation based on isolated fractions at specified pI ranges, closely matched migration time and baseline-resolved peak shape makes the device a general tool to detect unknown proteins and determine known minor proteins in protein-rich samples with interfering constituents.
Persistent Identifierhttp://hdl.handle.net/10722/203609
ISSN
2015 Impact Factor: 3.926
2015 SCImago Journal Rankings: 1.774
ISI Accession Number ID

 

DC FieldValueLanguage
dc.contributor.authorWu, Ren_US
dc.contributor.authorWang, Zen_US
dc.contributor.authorZhao, Wen_US
dc.contributor.authorYeung, WSBen_US
dc.contributor.authorFung, YSen_US
dc.date.accessioned2014-09-19T15:32:35Z-
dc.date.available2014-09-19T15:32:35Z-
dc.date.issued2013en_US
dc.identifier.citationJournal of Chromatography A, 2013, v. 1304, p. 220-226en_US
dc.identifier.issn0021-9673-
dc.identifier.urihttp://hdl.handle.net/10722/203609-
dc.description.abstractTo improve resolution of important minor proteins and eliminate time-consuming precipitation of major protein with associated analyte co-precipitation risk, a multi-dimension strategy is adopted in the 2D microchip-CE device to isolate major proteins on-chip, enrich minor proteins in capillary before their separation in CE for UV quantitation. A standard fluorescent protein mixture containing FITC-BSA, myoglobin and cytochrome as specific pI markers has prepared to demonstrate capability of the device to fractionate minor proteins by IEF. The results using a standard protein mixture with profile resembling infant milk formula show a complete isolation of high abundance proteins by a 2-min 1D IEF run. The subsequent t-ITP/CZE run by on-chip high voltage switching delivers a high stacking ratio, realizing 60 folds enrichment of isolated protein fractions. All five important functional proteins (LF, IgG, alpha-LA, beta-LgA and beta-LgB) known to fortify infant milk formula are isolated and determined using two consecutive t-ITP-CZE runs within a 18-min total assay time, a significant saving compared to several hours conventional pretreatment. For a 100g infant milk formula sample, working ranges of 20-8000mg, repeatability 3.8-5.3% and detection limits 2.3-10mg have been achieved to meet government regulations. Method reliability is established by 100% recoveries and agreeable results within expected ranges and labeled values. The capability of the device for field operation, rapid assay with quick results, label-free universal detection, simple operation by aqueous dissolution before injection, and the demanding matching in 2D separation based on isolated fractions at specified pI ranges, closely matched migration time and baseline-resolved peak shape makes the device a general tool to detect unknown proteins and determine known minor proteins in protein-rich samples with interfering constituents.-
dc.languageengen_US
dc.publisherElsevier BV. The Journal's web site is located at http://www.elsevier.com/locate/chroma-
dc.relation.ispartofJournal of Chromatography Aen_US
dc.subject.meshElectrophoresis, Microchip - economics - instrumentation-
dc.subject.meshInfant Formula - chemistry-
dc.subject.meshMilk Proteins - isolation and purification-
dc.subject.meshReproducibility of Results-
dc.subject.meshSerum Albumin, Bovine - isolation and purification-
dc.titleMulti-dimension microchip-capillary electrophoresis device for determination of functional proteins in infant milk formula.en_US
dc.typeArticleen_US
dc.identifier.emailZhao, W: wfzhao@hku.hken_US
dc.identifier.emailYeung, WSB: wsbyeung@hkucc.hku.hk-
dc.identifier.emailFung, YS: ysfung@hku.hk-
dc.identifier.doi10.1016/j.chroma.2013.06.073-
dc.identifier.pmid23870546-
dc.identifier.scopuseid_2-s2.0-84881028297-
dc.identifier.hkuros238665en_US
dc.identifier.hkuros240985-
dc.identifier.volume1304en_US
dc.identifier.spage220en_US
dc.identifier.epage226en_US
dc.identifier.isiWOS:000323017500027-
dc.publisher.placeNetherlands-

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