Distance-Based Tear Lactoferrin Assay on Microfluidic Paper Device Using Interfacial Interactions on Surface-Modified Cellulose

Kentaro Yamada, Terence G. Henares, Koji Suzuki, Daniel Citterio

Research output: Contribution to journalArticle

60 Citations (Scopus)

Abstract

Distance-based detection motifs on microfluidic paper-based analytical devices (μPADs) allow quantitative analysis without using signal readout instruments in a similar manner to classical analogue thermometers. To realize a cost-effective and calibration-free distance-based assay of lactoferrin in human tear fluid on a μPAD not relying on antibodies or enzymes, we investigated the fluidic mobilities of the target protein and Tb3+ cations used as the fluorescent detection reagent on surface-modified cellulosic filter papers. Chromatographic elution experiments in a tear-like sample matrix containing electrolytes and proteins revealed a collapse of attractive electrostatic interactions between lactoferrin or Tb3+ and the cellulosic substrate, which was overcome by the modification of the paper surface with the sulfated polysaccharide I-carrageenan. The resulting μPAD based on the fluorescence emission distance successfully analyzed 0-4 mg mL-1 of lactoferrin in complex human tear matrix with a lower limit of detection of 0.1 mg mL-1 by simple visual inspection. Assay results of 18 human tear samples including ocular disease patients and healthy volunteers showed good correlation to the reference ELISA method with a slope of 0.997 and a regression coefficient of 0.948. The distance-based quantitative signal and the good batch-to-batch fabrication reproducibility relying on printing methods enable quantitative analysis by simply reading out concentration scale marks printed on the μPAD without performing any calibration and using any signal readout instrument.

Original languageEnglish
Pages (from-to)24864-24875
Number of pages12
JournalACS Applied Materials and Interfaces
Volume7
Issue number44
DOIs
Publication statusPublished - 2015 Nov 11

Keywords

  • anionic polysaccharide
  • cation immobilization
  • paper-based analytical device
  • protein mobility
  • surface modification

ASJC Scopus subject areas

  • Materials Science(all)

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