title: Screening protein isoforms predictive for cancer using immunoaffinity capture and fast LC-MS in PRM mode.
abstract: OBJECTIVE
  We report an immunocapture strategy to extract proteins known to harbor driver mutations for a defined cancer type before the simultaneous assessment of their mutational status by MS. Such a method bypasses the sensitivity and selectivity issues encountered during the analysis of unfractionated complex biological samples.
  
  
  METHODS
  Fast LC separations using short nanobore columns hyphenated with a high-resolution quadrupole-orbitrap mass spectrometer have been devised to take advantage of fast MS cycle times in conjunction with sharp chromatographic peak widths to accelerate the sample analysis throughput. Such an analytical platform is well suited to analyze simple protein mixtures obtained after immunoaffinity enrichment.
  
  
  RESULTS
  After establishing the technical performance of the platform, the method was applied to the quantitative profiling of cellular Ras and EGFR protein isoforms, as well as serum amyloid A isoforms in plasma.
  
  
  CONCLUSIONS
  Immunoaffinity purification combined with fast LC-MS detection for the detection of driver mutations in tissue and tumor biomarkers in plasma samples can assist clinicians to select an optimal therapeutic intervention for patients.
authors: Luxembourg Clinical Proteomics Center (LCP), CRP-Santé, Strassen, Luxembourg.
authors: Antoine
authors: A
authors: Lesur
authors: Luxembourg Clinical Proteomics Center (LCP), CRP-Santé, Strassen, Luxembourg.
authors: Lina
authors: L
authors: Ancheva
authors: Luxembourg Clinical Proteomics Center (LCP), CRP-Santé, Strassen, Luxembourg.
authors: Yeoun Jin
authors: YJ
authors: Kim
authors: Laboratory of Experimental Hemato-Oncology, CRP-Santé, Strassen, Luxembourg.|Centre Hospitalier Luxembourg (CHL), Strassen, Luxembourg.
authors: Guy
authors: G
authors: Berchem
authors: Luxembourg Clinical Proteomics Center (LCP), CRP-Santé, Strassen, Luxembourg.
authors: Jan
authors: J
authors: van Oostrum
authors: Luxembourg Clinical Proteomics Center (LCP), CRP-Santé, Strassen, Luxembourg.
authors: Bruno
authors: B
authors: Domon
mesh_terms: D000595:Amino Acid Sequence; D045744:Cell Line, Tumor; D002846:Chromatography, Affinity / Q000379:methods*; D002853:Chromatography, Liquid / Q000379:methods*; D006801:Humans; D013058:Mass Spectrometry / Q000379:methods*; D008969:Molecular Sequence Data; D009154:Mutation / Q000235:genetics; D009363:Neoplasm Proteins / Q000097:blood / Q000378:metabolism; D009369:Neoplasms / Q000097:blood / Q000378:metabolism*; D010455:Peptides / Q000737:chemistry; D020033:Protein Isoforms / Q000097:blood / Q000378:metabolism*; D016415:Sequence Alignment; D018631:ras Proteins / Q000378:metabolism
chemical_list: D009363:Neoplasm Proteins; D010455:Peptides; D020033:Protein Isoforms; D018631:ras Proteins
keywords: EGFR; Fast LC; KRas; PLASMA; PRM; SAA