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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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- TARGETED ANALYSIS -

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Targeted analysis

sHPP

- TARGETED ANALYSIS -

Adán Alpízar Morúa

Proteomics Unit. Centro Nacional de Biotecnología


Targeted analysis

  • AssignedProteins


Targeted analysis

  • Methods (MRM Pilot)

1


Targeted analysis

AEAESR

331.7/462.2

331.7/591.3

331.7/488.2

331.7/401.2


Targeted analysis

GPLEYPSAK

530.8/1003.6

530.8/664.4

530.8/793.4

530.8/906.5


Targeted analysis

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


Targeted analysis

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


Targeted analysis

3

  • PeptideSynthesis


Targeted analysis

3

  • PeptideSynthesis (MRM)


Targeted analysis

  • 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


Targeted analysis

  • acknowledgments

Fernando Roncal

Manuel Lombardía

Miguel Marcilla

SeverineGharbi

Carmen González

RosanaNavajas

Alberto Paradela

Salvador Martínez

Alberto Medina


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