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High-Throughput Analysis of Proteins in CCD18 and MCF7/RAMOS Cell Lines Using LC-MS

This study, conducted by Marta Mendes and Ignacio Casal at the Centro de Investigaciones Biológicas in August 2012, explores the protein profiles of CCD18 and MCF7/RAMOS cell lines. Using high-throughput LC-MS techniques, we analyzed protein digests obtained through trypsin digestion under stringent conditions with a focus on proteotypic peptides. The investigation utilized state-of-the-art LTQ Orbitrap Velos equipment and implemented advanced data analysis using MaxQuant and Proteome Discoverer. Our findings identify isoforms and potential biomarkers relevant to cancer research.

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High-Throughput Analysis of Proteins in CCD18 and MCF7/RAMOS Cell Lines Using LC-MS

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  1. Chromosome 16 Marta Mendes Ignacio Casal Centro de Investigaciones Biológicas August 2012

  2. 250 CCD18 150 Proteins to search for 100 75 50 37 25 20 30 mg/lane 15 10 Cell Lines CCD18 MCF7 Ramos 1D SDS PAGE MCF7 RAMOS 250 150 100 75 250 150 50 100 75 37 50 30 mg/lane 30 mg/lane 37 25 20 High throughput analysis 25 Digestion with trypsin 20 15 15 10 10

  3. Equipment : LTQ Orbitrap Velos coupled to a Proxeon nanoLC system LC conditions MS System

  4. High throughput DDA Top15 Proteome discoverer and Sequest with FDR 1% Database: uniprot_sprot_human_HPP KLEEENENLR CCD18 No proteins found MCF7 Q8TBY8-2 RAMOS

  5. Q8TBY8 isoforms alignment

  6. Target using global mass list Proteotypic peptides obtained at SRM Atlas LC conditions MS System MAXQUANT Database: uniprot_sprot_human_HPP

  7. EFQEEMAALK LMEGELEALR

  8. Q8TBY8 isoforms alignment Q8TBY8 isoforms 1 and 2 are present in both CCD18 and Ramos cell lines

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