Detection of Mast Cells Expressing c-Kit Using Antibody Covalently Bound to Gelatin Elongated from Surface of Immunosensor Based on Surface Plasmon Resonance

Anal Sci. 2019 Jul 10;35(7):811-813. doi: 10.2116/analsci.19N012. Epub 2019 Mar 29.

Abstract

An immunosensor based on surface plasmon resonance was applied to detect mast cells expressing c-Kit. Sufficient detection of the mast cells was achieved by covalent immobilization of gelatin firstly on the sensor surface and followed by covalent binding of the anti-c-Kit antibody to lysine residues in the gelatin molecules through bis(sulfosuccinimidyl)suberate (BS3) treatment. By using BS3, which is a homo-bifunctional reagent, the lysine residues of the anti-c-Kit antibody easily bound to the lysine residues of the gelatin in the physiological condition. The lower limit of detection was 104 cells/mL.

Keywords: SPR; antibody; bone marrow-derived mast cells; c-Kit; gelatin; immunosensor.

MeSH terms

  • Animals
  • Antibodies, Immobilized / chemistry*
  • Antibodies, Immobilized / immunology
  • Decanoic Acids / chemistry
  • Gelatin / chemistry*
  • Gene Expression Regulation*
  • Humans
  • Immunoassay / methods*
  • Limit of Detection
  • Mast Cells / cytology*
  • Mast Cells / metabolism
  • Mice
  • Proto-Oncogene Proteins c-kit / immunology
  • Proto-Oncogene Proteins c-kit / metabolism*
  • Succinimides / chemistry
  • Surface Plasmon Resonance / methods*

Substances

  • Antibodies, Immobilized
  • Decanoic Acids
  • Succinimides
  • bis(sulfosuccinimidyl) sebacate
  • Gelatin
  • Proto-Oncogene Proteins c-kit