Characterization of binding and quantification of human autoantibodies to PDGFRα using a biosensor-based approach

Anal Biochem. 2017 Jul 1;528:26-33. doi: 10.1016/j.ab.2017.04.011. Epub 2017 Apr 24.

Abstract

Systemic sclerosis (SSc) is a chronic autoimmune disease of the connective tissue. The variety and clinical relevance of autoantibodies in SSc patients have been extensively studied, eventually identifying agonistic autoantibodies targeting the platelet-derived growth factor receptor alpha (PDGFRα), and representing potential biomarkers for SSc. We used a resonant mirror biosensor to characterize the binding between surface-blocked PDGFRα and PDGFRα-specific recombinant human monoclonal autoantibodies (mAbs) produced by SSc B cells, and detect/quantify serum autoimmune IgG with binding characteristics similar to the mAbs. Kinetic data showed a conformation-specific, high-affinity interaction between PDGFRα and mAbs, with equilibrium dissociation constants in the low-to-high nanomolar range. When applied to total serum IgG, the assay discriminated between SSc patients and healthy controls, and allowed the rapid quantification of autoimmune IgG in the sera of SSc patients, with anti-PDGFRα IgG falling in the range 3.20-4.67 neq/L of SSc autoantibodies. The test was validated by comparison to direct and competitive anti-PDGFRα antibody ELISA. This biosensor assay showed higher sensibility with respect to ELISA, and other major advantages such as the specificity, rapidity, and reusability of the capturing surface, thus representing a feasible approach for the detection and quantification of high affinity, likely agonistic, SSc-specific anti-PDGFRα autoantibodies.

Keywords: Autoantibody quantification; Binding kinetics; Biosensor; PDGFR; Systemic sclerosis.

Publication types

  • Evaluation Study

MeSH terms

  • Adult
  • Aged
  • Autoantibodies / blood*
  • Autoantibodies / immunology*
  • B-Lymphocytes / immunology
  • Biomarkers / blood*
  • Biosensing Techniques / methods*
  • Female
  • Humans
  • Limit of Detection
  • Male
  • Middle Aged
  • Receptor, Platelet-Derived Growth Factor alpha / immunology*
  • Scleroderma, Systemic / diagnosis
  • Scleroderma, Systemic / immunology*
  • Sensitivity and Specificity

Substances

  • Autoantibodies
  • Biomarkers
  • Receptor, Platelet-Derived Growth Factor alpha