Quantifying antibody binding on protein microarrays using microarray nonlinear calibration

Biotechniques. 2013 May;54(5):257-64. doi: 10.2144/000114028.


We present a microarray nonlinear calibration (MiNC) method for quantifying antibody binding to the surface of protein microarrays that significantly increases the linear dynamic range and reduces assay variation compared with traditional approaches. A serological analysis of guinea pig Mycobacterium tuberculosis models showed that a larger number of putative antigen targets were identified with MiNC, which is consistent with the improved assay performance of protein microarrays. MiNC has the potential to be employed in biomedical research using multiplex antibody assays that need quantitation, including the discovery of antibody biomarkers, clinical diagnostics with multi-antibody signatures, and construction of immune mathematical models.

Publication types

  • Research Support, N.I.H., Extramural

MeSH terms

  • Algorithms
  • Animals
  • Antigens, Bacterial / chemistry
  • Antigens, Bacterial / immunology
  • Antigens, Bacterial / metabolism
  • Biomarkers / analysis
  • Biomarkers / blood
  • Biomarkers / metabolism
  • Calibration
  • Guinea Pigs
  • Humans
  • Immunoglobulin G / analysis*
  • Immunoglobulin G / blood
  • Immunoglobulin G / immunology
  • Immunoglobulin G / metabolism
  • Mice
  • Mycobacterium tuberculosis / immunology
  • Nonlinear Dynamics
  • Protein Array Analysis / methods*
  • Protein Binding
  • Reference Standards
  • Tuberculosis, Pulmonary / immunology
  • Tumor Suppressor Protein p53 / chemistry
  • Tumor Suppressor Protein p53 / immunology
  • Tumor Suppressor Protein p53 / metabolism


  • Antigens, Bacterial
  • Biomarkers
  • Immunoglobulin G
  • Tumor Suppressor Protein p53