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ZÍTKA, O. SKALIČKOVÁ, S. KŘÍŽKOVÁ, S. VLKOVÁ, M. ADAM, V. KIZEK, R.
Originální název
Separation of Lactoferrin from Human Saliva Using Monolithic Disc
Typ
článek v časopise ve Web of Science, Jimp
Jazyk
angličtina
Originální abstrakt
In this study, we optimized method for the isolation and detection of lactoferrin from human saliva using 3 mm short monolithic disc. We optimized the conditions for separation as flow rate 4 mL min(-1) and ionic strength of effluent as 2 M center dot NaCl. We estimated limit of detection of whole method, which was hyphenated to the Bradford's assay, down to 100 ng mL(-1). The purity of the isolated fractions was verified by sodium dodecyl sulphate-polyacrylamide gel electrophoresis and recovery of isolation was found to be 51 % using minimally processed sample of saliva. Further, we tested the optimized method on group of healthy volunteers (n = 7). We were able to distinguish between the healthy subjects and subject suffering from celiac disease, which reported at least 2.5x higher level of lactoferrin in comparison to healthy ones. The results were correlated with standard enzyme-linked immunosorbent assay (ELISA) kit with obtained correlation coefficient R-2 = 0.8446. Analysis of lactoferrin in saliva by monolithic disc and subsequent offline photometric detection is faster and cheaper method compared to ELISA commercial kit. The total analysis of one sample takes <20 min.
Klíčová slova
Liquid chromatography; Monolithic disc; Spectrophotometry; Enzyme-linked immunosorbent assay; Immunodiagnostics; Human saliva; Lactoferrin
Autoři
ZÍTKA, O.; SKALIČKOVÁ, S.; KŘÍŽKOVÁ, S.; VLKOVÁ, M.; ADAM, V.; KIZEK, R.
Vydáno
1. 6. 2013
ISSN
0009-5893
Periodikum
CHROMATOGRAPHIA
Ročník
76
Číslo
11-12
Stát
Spolková republika Německo
Strany od
611
Strany do
619
Strany počet
9
BibTex
@article{BUT145218, author="Ondřej {Zítka} and Sylvie {Skaličková} and Soňa {Křížková} and Marcela {Vlková} and Vojtěch {Adam} and René {Kizek}", title="Separation of Lactoferrin from Human Saliva Using Monolithic Disc", journal="CHROMATOGRAPHIA", year="2013", volume="76", number="11-12", pages="611--619", doi="10.1007/s10337-013-2459-x", issn="0009-5893" }