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sHPP. - TARGETED ANALYSIS -. Adán Alpízar Morúa. Proteomics Unit. Centro Nacional de Biotecnología. Assigned Proteins. Methods (MRM Pilot ). 1. AEAESR. 331.7/462.2. 331.7/591.3. 331.7/488.2. 331.7/401.2. GPLEYPSAK. 530.8/1003.6. 530.8/664.4. 530.8/793.4. 530.8/906.5. 2.
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sHPP - TARGETED ANALYSIS - Adán Alpízar Morúa Proteomics Unit. Centro Nacional de Biotecnología
AEAESR 331.7/462.2 331.7/591.3 331.7/488.2 331.7/401.2
GPLEYPSAK 530.8/1003.6 530.8/664.4 530.8/793.4 530.8/906.5
2 QITQVYGFYDECLR (+2) EILVEESNVQR (+2) 896.4/855.4 685.3/861.4 • MCF7 (SDS/PAGE Band 8) P60510 896.4/1059.5 685.3/960.5 896.4/1222.5 685.3/1073.6 QITQVYGFYDECLRK (+3) 640.6/983.5 640.6/1130.5 640.6/1187.5
3 • PeptideSynthesis Selection Criteria of the Synthetic Peptides • 3-4 peptides per protein • Unique peptides • Peptide Length: 9-21 residues • No missed cleavages • No Mand W • No Qby N-ter • MRM atlas
3 • PeptideSynthesis
3 • PeptideSynthesis (MRM)
Conclusions Currently, data depedent analysis yields better results in terms of sensitivity and number of identifications than MRM We have not yet implemented a robust workflow for the routine implementation of MRM methods We have implemented a platform that allows the simultaneous synthesis of a significant (≈ 60) number of peptides
acknowledgments Fernando Roncal Manuel Lombardía Miguel Marcilla SeverineGharbi Carmen González RosanaNavajas Alberto Paradela Salvador Martínez Alberto Medina