Analytical and Bioanalytical Chemistry

, Volume 408, Issue 5, pp 1497–1506 | Cite as

Anti-heat shock protein autoantibody profiling in breast cancer using customized protein microarray

  • Liu Shi
  • Thomas Gehin
  • Yann Chevolot
  • Eliane Souteyrand
  • Alain Mangé
  • Jérôme Solassol
  • Emmanuelle LaurenceauEmail author
Research Paper


Heat shock proteins (HSPs) are over-expressed in a wide range of human cancers. It results in the stimulation of the immune system and consequently in elevated concentration of anti-HSP autoantibodies. Elevated anti-HSP autoantibodies were found in breast cancer patients, and they are associated with tumor metastasis. Therefore, screening these autoantibodies could be of diagnostic and prognostic values. Protein microarrays have already demonstrated their great potential as a diagnostic tool. However, protein diversity requires optimization of the microarray fabrication to achieve high sensitivity and specificity. In this study, seven HSPs were immobilized on six different surface chemistries. After evaluation and optimization with purified antibodies of the six surface chemistries, two surfaces were selected to detect anti-HSP autoantibodies in breast cancer sera. Multiplex detection of anti-HSP autoantibodies allowed discrimination of breast cancer patients (50) from healthy controls (26) with a sensitivity of 86 % and a specificity of 100 %.

Graphical abstract

Receiver operating characteristic (ROC) curve analysis for the discrimination between breast cancer patients and healthy controls: detection of individual autoantibody on optimal surface chemistry (COOH or chitosan surface) and combination (black line) of the detection of 7 autoantibodies


Breast cancer Autoantibodies Heat shock proteins Protein microarray 



China Scholarship Council is acknowledged for offering a Ph.D. fellowship to Liu Shi. The CNANO Rhône-Alpes and BQR Ecole Centrale de Lyon are acknowledged for financial support. NanoLyon is acknowledged for technical support.

Compliance with ethical standards

Conflict of interest

The authors declare that they have no competing interests.

Informed consent

All human samples were prospectively collected between 2005 and 2007 at the CRLC Val d’Aurelle Cancer Institute (Montpellier, France) at the time of cancer diagnosis after obtaining written informed consent. This study was approved by the Montpellier University Hospital human research committee and the INSERM review board (RBM-03-63).

Supplementary material

216_2015_9257_MOESM1_ESM.pdf (252 kb)
ESM 1 (PDF 252 kb)


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Copyright information

© Springer-Verlag Berlin Heidelberg 2015

Authors and Affiliations

  • Liu Shi
    • 1
  • Thomas Gehin
    • 1
  • Yann Chevolot
    • 1
  • Eliane Souteyrand
    • 1
  • Alain Mangé
    • 2
  • Jérôme Solassol
    • 2
  • Emmanuelle Laurenceau
    • 1
    Email author
  1. 1.Institut des Nanotechnologies de Lyon INL-UMR5270, CNRS, Ecole Centrale de LyonUniversité de LyonEcullyFrance
  2. 2.Institut de Recherche en Cancérologie de Montpellier INSERM U1194MontpellierFrance

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