Publication detail

Separation of Lactoferrin from Human Saliva Using Monolithic Disc

ZÍTKA, O. SKALIČKOVÁ, S. KŘÍŽKOVÁ, S. VLKOVÁ, M. ADAM, V. KIZEK, R.

Original Title

Separation of Lactoferrin from Human Saliva Using Monolithic Disc

Type

journal article in Web of Science

Language

English

Original Abstract

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.

Keywords

Liquid chromatography; Monolithic disc; Spectrophotometry; Enzyme-linked immunosorbent assay; Immunodiagnostics; Human saliva; Lactoferrin

Authors

ZÍTKA, O.; SKALIČKOVÁ, S.; KŘÍŽKOVÁ, S.; VLKOVÁ, M.; ADAM, V.; KIZEK, R.

Released

1. 6. 2013

ISBN

0009-5893

Periodical

CHROMATOGRAPHIA

Year of study

76

Number

11-12

State

Federal Republic of Germany

Pages from

611

Pages to

619

Pages count

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"
}