ScFv-modified graphene-coated IDE-arrays for 'label-free' screening of cardiovascular disease biomarkers in physiological saline

Biosens Bioelectron. 2018 Apr 15:102:574-581. doi: 10.1016/j.bios.2017.12.005. Epub 2017 Dec 7.

Abstract

Fatty-acid binding proteins (FABP) and myeloperoxidases (MPO) are associated with many chronic conditions in humans and considered to be important biomarkers for diagnosis of cardiac diseases. Here we assemble a new electrical biosensor platform based on graphene-coated interdigitated electrode arrays (IDE-arrays) towards ultrafast, label-free screening of heart type-FABP and MPO. Arrays of nanoscale (nanoIDE) and microscale (microIDE) electrode-arrays were fabricated on wafer-scale by combining nanoimprint and photolithography processes. Chemical vapor deposition grown multilayer graphene was transferred onto nano/microIDE-arrays and used as a high surface-to-volume ratio electrical transducer. Novel biofunctional layers of specially engineered anti-h-FABP and anti-MPO single-chain fragment variables (scFv) were immobilized onto graphene-coated IDE-array sensor platform for electrical detection of h-FABP and MPO in physiological saline. scFv fragments show increased sensitivity in comparison to the state-of-the-art competitive ELISA for their higher affinity towards target analytes. Deploying FABP and MPO specific scFvs as receptor molecules onto our high-sensitivity graphene-coated IDE-arrays with identical sensor characteristics and assays covering clinically relevant concentrations in physiological saline, we demonstrate realization of a simple and versatile biosensor platform capable of high performance cardiac-bioassays for point-of-care applications.

Keywords: Biosensor; Cardiovascular; Electrical sensing; Graphene; Physiological buffer; scFV.

MeSH terms

  • Biomarkers / blood
  • Biosensing Techniques / methods*
  • Cardiovascular Diseases / blood*
  • Fatty Acid Binding Protein 3 / blood
  • Fatty Acid Binding Protein 3 / immunology
  • Fatty Acid Binding Protein 3 / isolation & purification*
  • Granulocyte Colony-Stimulating Factor / blood
  • Granulocyte Colony-Stimulating Factor / immunology
  • Granulocyte Colony-Stimulating Factor / isolation & purification*
  • Graphite / chemistry
  • Humans
  • Interleukin-3 / blood
  • Interleukin-3 / immunology
  • Interleukin-3 / isolation & purification*
  • Recombinant Fusion Proteins / blood
  • Recombinant Fusion Proteins / immunology
  • Recombinant Fusion Proteins / isolation & purification*
  • Single-Chain Antibodies / chemistry
  • Single-Chain Antibodies / immunology

Substances

  • Biomarkers
  • Fatty Acid Binding Protein 3
  • Interleukin-3
  • Recombinant Fusion Proteins
  • Single-Chain Antibodies
  • myelopoietin
  • Granulocyte Colony-Stimulating Factor
  • Graphite