Acetonitril

Alkaline phosphatase (AP)-conjugated goat-anti-human IgG, streptavidin, AP substrate tablets (para-nitrophenylphosphate (pNPP)), Tris, Tween, sodium chloride, sodium carbonate, ethanolamine, MgCl₂, D-Biotin, hexafluorophosphate azabenzotriazole tetramethyl uranium (HATU), milliQ water, trifluoroacetic acid (TFA), triisopropylsilane (TIS), and N,N-diisopropylethylamine (DIPEA) were from Sigma Aldrich (St. Louis, MO, USA). Synthetic EBNA1 and EBNA2 peptides were purchased from Schäfer-N (Lyngby, Denmark) (Table 1), and were generated on TentaGel resin using standard Fmoc-based solid-phase peptide synthesis (SPPS) [54 (link)]. The peptides were synthesized as peptide acids. EBNA3 peptides were synthesized by microwave-assisted SPPS [55 (link),56 (link)]. Wang resin, Oxyma Pure, and DIC was from Iris Biotech AG (Marktredwitz, Germany). Fmoc-L-amino acids were from Novabiochem (Darmstadt, Germany). Acetonitril was from Carlo Erba (Milano, Italia).

dithiothreitol (DTT), pronase, ethylenediamine tetra-acetic acid (EDTA), collagen IV, urea, thio-urea, orthovanadate, glycerol, TRIS, iodoacetamide, NaCl, detergents (CHAPS, sodium deoxychotate, Triton X100), NH4CO3 and the Glu-fibrinopeptide were from Sigma-Aldrich (Saint-Quentin Fallavier, France). Anti-proteases were from Roche (Neuilly sur Seine, France). Bromophenol blue, SDS, acrylamide, silver staining kit were from Biorad (Marnes la Coquette, France). Trypsin for mass spectrometry was of highest grade purchased from Promega (Charbonnieres-les-bains, France). Acetonitril (ACN) and Tri-fluoro-acetic acid (TFA) were from Carlo Erba (Val de Reuil, France), Alpha-Cyano-4-Hydroxycinnamic Acid (HCCA) from Laser Biolabs (Sophia-Antipolis, France). iTRAQ reagents were from Applied Biosystems (Courtaboeuf, France)