mRNA PCR-Based Epitope Chase Method

  • Jean-Daniel Doucet
  • Dominique Gauchat
  • Réjean Lapointe
Part of the Methods in Molecular Biology book series (MIMB, volume 969)


The identification of specific viral and tumor antigen epitopes recognized by CD4+ or CD8+ T lymphocytes remains a challenge. Unfortunately, epitope mapping methods are generally costly and time-consuming. This chapter details a polymerase chain reaction (PCR)-based mRNA epitope identification method called mPEC, which is designed to rapidly and precisely identify relevant T cell epitopes recognized by previously isolated CD8+ or CD4+ T lymphocytes.

This method is based on the use of mRNA fragments synthesized from PCR-amplified cDNA with a variety of 3′end iterative deletions. mRNA fragments are electroporated into autologous antigen-presenting cells to map the epitope in a given protein antigen. Considering mRNA’s sensitivity to degradation, we also insert a control define epitope at the mRNA’s 3′end to control for electroporated mRNA’s integrity and capacity to be translated.

Key words

Major histocompatibility complexes class I and II T cell epitope Epitope mapping mRNA transfection 



This work was supported by the Canadian Institutes of Health Research (CIHR). R.L. and J.-D.D. are recipients of scholarships from the Fonds de la recherche en santé du Québec (FRSQ). The authors would like to thank K. Lieber and L.-A. Hanafi for text editing.


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Copyright information

© Springer Science+Business Media New York 2013

Authors and Affiliations

  • Jean-Daniel Doucet
    • 1
  • Dominique Gauchat
    • 1
  • Réjean Lapointe
    • 1
  1. 1.Research Centre, Centre Hospitalier de l’Université de Montréal (CRCHUM), Hôpital Notre-Dame, Université de Montréal and Institut du Cancer de Montréal (ICM)MontrealCanada

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