Please use this identifier to cite or link to this item: http://hdl.handle.net/11455/67884
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dc.contributor.authorHo, J.A.A.en_US
dc.contributor.authorHung, C.H.en_US
dc.contributor.authorWu, L.C.en_US
dc.contributor.authorLiao, M.Y.en_US
dc.date2009zh_TW
dc.date.accessioned2014-06-11T05:55:53Z-
dc.date.available2014-06-11T05:55:53Z-
dc.identifier.issn0003-2700zh_TW
dc.identifier.urihttp://hdl.handle.net/11455/67884-
dc.description.abstractA folic acid-anchored, poly(ethylene glycol)-linked (PEGgylated) phospholipid and an immunoaffinity chromatographic column were prepared and employed to develop a liposomal immunodiagnostic assay for the direct determination of folic acid (FA) in this study. Distearoylphosphatidylethanolamine-poly(ethylene glycol)(2000)-folic acid (DSPE-PEG(2000)-FA) was synthesized through carbodiimide-mediated coupling of FA and DSPE-PEG(2000)-amine and characterized using thin layer chromatography, I H nuclear magnetic resonance spectroscopy, and electrospray ionization-mass spectrometry. Liposomal biolabels were constructed using the synthesized DSPE-PEG(2000)-FA in conjunction with other phospholipids. A stationary phase having affinity for FA was prepared by covalently linking purified anti-FA monoclonal antibodies onto N-hydroxysuccinimide-activated Sepharose beads, which were subsequently packed into a 1.9 cm diameter polypropylene column. The calibration curve for FA had a linear range from 10(-8) to 10(-4) M. The limit of detection was 6.8 ng (equivalent to 500 mu L of 3.1 x 10(-8) M FA). The elution buffer (35% methanol in Tris buffered saline containing 0.1% Tween 20) also served as the regeneration buffer, which allowed the same column to be used for up to 50 times without any observable loss of reactivity. The immunoaffinity chromatographic column was reusable and capable of concentrating analytes from sample solution; in conjunction with folic acid-sensitized liposomal biolabels, however, they hold great potential as sensitive immunoaffinity assays for the determination for FA. To confirm the feasibility of using this system in the analysis of real samples, the folic acid contents of three over-the-counter vitamin supplements were tested. The recoveries of folic acid of 90-112% for these three samples were obtained, suggesting contents that were consistent with the information obtained from their nutritional facts panels.en_US
dc.language.isoen_USzh_TW
dc.relationAnalytical Chemistryen_US
dc.relation.ispartofseriesAnalytical Chemistry, Volume 81, Issue 14, Page(s) 5671-5677.en_US
dc.relation.urihttp://dx.doi.org/10.1021/ac900402ven_US
dc.subjectperformance liquid-chromatographyen_US
dc.subject96-well microtiter platesen_US
dc.subjectmicrobiological assayen_US
dc.subjectvascular-diseaseen_US
dc.subjectfolate receptoren_US
dc.subjectin-vitroen_US
dc.subjectfooden_US
dc.subjectmilken_US
dc.subjecthomocysteineen_US
dc.subjectimmunoassayen_US
dc.titleFolic Acid-Anchored PEGgylated Phospholipid Bioconjugate and Its Application in a Liposomal Immunodiagnostic Assay for Folic Aciden_US
dc.typeJournal Articlezh_TW
dc.identifier.doi10.1021/ac900402vzh_TW
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