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Proceedings of the 31 st European Peptide SymposiumMichal Lebl, Morten Meldal, Knud J. Jensen, Thomas Hoeg-Jensen (Editors)European Peptide Society, 2010NCeliac Disease: Characterization of Amino Acids and ShortPeptides Finger Print for a Fast I.V.D.Margherita Di Pisa 1,2 , Giuseppina Sabatino 1,3 , Mario Chelli 1,2 ,Paolo Rovero 1,5 , Claudio Tiberti 6 , and Anna Maria Papini 1,2,41 Laboratory of Peptide & Protein Chemistry & Biology, University of Florence, I-50019,Sesto Fiorentino, Italy; 2 Department of Chemistry “Ugo Schiff” and CNR-ICCOM,University of Florence, I-50019, Sesto Fiorentino, Italy; 3 Espikem Srl, I-59100, Prato,Italy; 4 Laboratoire SOSCO – EA4505 Université de Cergy-Pontoise, Neuville-sur-Oise,F-95031, Cergy-Pontoise, France; 5 Department of Pharmaceutical Sciences, University ofFlorence, I-50019, Sesto Fiorentino, Italy; 6 Department of Clinical Sciences, PoliclinicoUmberto I, Sapienza, University of Rome, ItalyIntroductionCeliac Disease (CD) is an autoimmune disease characterized by villous atrophy andinflammatory cell infiltration of the lamina propria caused by the gliadin fraction of wheatgluten. The endomysial protein tissue Transglutaminase (tTG) is the main autoantigen inCD playing a key role in the pathogenesis [1]. tTG catalyzes deamidation of Gln residuesor protein cross-linking through the formation of isopeptide bonds between Gln and Lysresidues in gliadin peptides that are rich in proline and glutamine and thus very resistant toproteolysis [2]. CD is characterized by the presence of specific antibodies recognizinggliadin, food proteins, and the endomysial autoantigen tissue Transglutaminase. Aminoacids and short peptides are important targets for metabolic profiling in many areas,including clinical diagnostics for evaluating disease conditions [3,4]. Total amino acidscontent in plasma and urine is expected to reflect the nutritional/metabolic status, i.e.confirming or ruling out the suspected aminoacidopathies. In order to achieve a betterunderstanding of this multifactorial disease, we are evaluating the free amino acids andshort peptides profile in serum of newly diagnosed celiac patients compared to healthysubjects by a quantitative AccQ•Tag Ultra UPLC analysis (Waters). Protocol analysis israther fast and includes a precolumn derivatization, which converts both primary andsecondary amino acids into exceptionally stable, fluorescent derivatives as suitable analytesfor UV-absorbance, fluorescence, electrochemical, and MS detection. Celiac patients’ sera,previously treated with sulfosalicilic acid to induce protein precipitation, were derivatizedwith a standard solution of 6-aminoquinolyl-N-hydroxysuccinimidyl carbamate (AQCreagent) (Figure 1).Results and discussionQuantification of individual amino acids has a relevant importance in monitoring thetherapeutic intervention by dietary manipulations and/or vitamin supplement therapy.A comparative evaluationNHNH20Ot1/2 ~ 15sNOOA Q C R e a g e n tNH2O6 - A m in o q u in o l i n e+NHR1R2Amino Acidt1/2