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Lactoferrin Isolation Using Monolithic Column Coupled with Spectrometric or Micro-Amperometric Detector.

Vojtech Adam1, Ondrej Zitka1, Petr Dolezal2

  • 1Department of Chemistry and Biochemistry, Mendel University of Agriculture and Forestry, Zemedelska 1, CZ-613 00 Brno, Czech Republic.

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|November 24, 2016
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Summary

This study developed an easy-to-use ion-exchange chromatography method for isolating lactoferrin from cow colostrum. The optimized technique achieved a low detection limit, paving the way for potential diagnostic applications.

Keywords:
ElectrochemistryLactoferrinLinear Sweep VoltammetryLiquid ChromatographyMilk ProteinMonolithic ColumnScreen-Printed Carbon ElectrodeUV-VIS Spectrometry

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Area of Science:

  • Biochemistry
  • Analytical Chemistry
  • Biotechnology

Background:

  • Lactoferrin is a protein with significant health benefits, including antimicrobial properties.
  • Accurate and efficient methods for lactoferrin isolation and quantification are crucial for its study and application.

Purpose of the Study:

  • To develop an easy-to-use technique for lactoferrin isolation from cow colostrum.
  • To optimize and validate separation and detection methods for lactoferrin analysis.
  • To explore miniaturization of detection systems for lactoferrin analysis in various samples.

Main Methods:

  • Sodium dodecyl sulphate-polyacrylamide gel electrophoresis (SDS-PAGE) and automated microfluidic electrophoresis were initially explored.
  • Ion-exchange chromatography (IEC) with a monolithic column and UV-VIS detection was optimized for lactoferrin isolation.
  • Amperometric detection using screen-printed carbon electrodes was investigated for miniaturized lactoferrin detection.

Main Results:

  • IEC with UV-VIS detection provided a detection limit of 0.1 μg/ml for lactoferrin in cow colostrum, with concentrations ranging from 0.5 to 1.1 g/l.
  • Miniaturized detection using self-fabricated screen-printed electrodes achieved a detection limit of 10 μg/ml, though commercial electrodes showed higher responses.
  • Lactoferrin levels in human faeces were below the detection limit of the miniaturized method in a healthy individual.

Conclusions:

  • Ion-exchange chromatography offers a viable and optimized method for routine lactoferrin analysis in cow colostrum.
  • Miniaturization efforts show promise for developing portable detection devices, though further optimization is needed.
  • Lactoferrin quantification in complex matrices like faeces requires highly sensitive detection methods.