Nano-yeast-scFv probes on screen-printed gold electrodes for detection of Entamoeba histolytica antigens in a biological matrix

Biosens Bioelectron. 2014 May 15:55:417-22. doi: 10.1016/j.bios.2013.12.043. Epub 2013 Dec 27.

Abstract

The time and costs associated with monoclonal antibody production limit the potential for portable diagnostic devices to penetrate the market. Replacing the antibody with a low-cost alternate affinity reagent would reduce the costs of diagnostic development and use, and lead to new portable diagnostic devices towards many diseases. Herein, we present low-cost affinity reagents, nano-yeast-scFv, on commercially available, inexpensive, and portable screen-printed electrodes for the label-free electrochemical detection of Entamoeba histolytica cyst antigens. The biosensor was able to detect antigen at concentrations down to 10 pg mL(-1) in buffer with an inter-assay reproducibility of (% RSD, n=3) 4.1%. The applicability of two differently engineered nano-yeast-scFv to each specifically detect their cognant E. histolytica cyst antigens was demonstrated in a biological matrix derived from human stool. Because of the simple, inexpensive, and sensitive nature of this methodology, it may offer a low-cost alternative to immunosensors based on antibody-target recognition.

Keywords: Affinity reagent; Electrochemical immunosensor; Entamoeba histolytica; Label-free detection; Nano-yeast–scFv; Screen-printed electrode.

Publication types

  • Research Support, N.I.H., Extramural
  • Research Support, Non-U.S. Gov't

MeSH terms

  • Antigens, Protozoan / analysis*
  • Antigens, Protozoan / immunology
  • Biosensing Techniques / instrumentation
  • Conductometry / instrumentation*
  • Electrodes*
  • Entamoeba histolytica / immunology*
  • Entamoeba histolytica / isolation & purification
  • Equipment Design
  • Equipment Failure Analysis
  • Feces / parasitology*
  • Fungal Proteins / immunology
  • Gold
  • Immunoassay / instrumentation*
  • Metal Nanoparticles / chemistry
  • Molecular Probe Techniques
  • Reproducibility of Results
  • Sensitivity and Specificity
  • Single-Chain Antibodies / immunology*

Substances

  • Antigens, Protozoan
  • Fungal Proteins
  • Single-Chain Antibodies
  • Gold