Electrochemical immunosensor based on gold-labeled monoclonal anti-LipL32 for leptospirosis diagnosis

Biosens Bioelectron. 2019 Oct 1;142:111539. doi: 10.1016/j.bios.2019.111539. Epub 2019 Jul 26.

Abstract

Leptospirosis is a critical human health problem in the tropical area, thus, a precise technique that can be used for point-of-care analysis is greatly required. This is the first report on electrochemical immunosensor based on gold-labeled monoclonal anti-LipL32 for rapid, simple and sensitive determination of LipL32. The sensor consisted of two LipL32-specific antibodies: an unlabeled capture primary antibody (Anti-1°Ab) and an electrochemically detectable gold-conjugated secondary antibody (Au-2°Ab). The Anti-1°Ab was immobilized onto the modified screen-printed graphene electrode (SPGE) to form the anti-LipL32 surface. The electrochemical signal response was determined by differential pulse voltammetry (DPV). In the presence of LipL32, the sensor displayed a significant increase in current response in a concentration-dependent manner, but no observable signal was detected in the absence of LipL32. The linearity between LipL32 concentration and the measured current was found in a range of 1-100 ng/mL, and the limit of detection (LOD) (3SDblank/Slope) and limit of quantitation (LOQ) (10SDblank/Slope) were found to be 0.28 and 0.93 ng/mL, respectively. This sensor was successfully applied to detect pathogenic Leptospira whole cell lysates samples with the satisfactory results. The promissing results suggested that this immunosensor might be an alternative tool for diagnosis of leptospirosis.

Keywords: Electrochemical immunosensor; Gold-labeled antibody; Graphene oxide; LipL32; Screen-printed graphene electrode.

MeSH terms

  • Antibodies, Immobilized / chemistry*
  • Antibodies, Monoclonal / chemistry
  • Bacterial Outer Membrane Proteins / analysis*
  • Biosensing Techniques / methods*
  • Electrochemical Techniques / methods
  • Gold / chemistry*
  • Graphite / chemistry
  • Humans
  • Immunoassay / methods
  • Leptospira / isolation & purification*
  • Leptospirosis / diagnosis
  • Limit of Detection

Substances

  • Antibodies, Immobilized
  • Antibodies, Monoclonal
  • Bacterial Outer Membrane Proteins
  • graphene oxide
  • Gold
  • Graphite