Proteomic Approaches by SELDI and MALDI-TOF/MS for CTL Analysis

  • Massimo PapaleEmail author
  • Maria Teresa Rocchetti
Part of the Methods in Molecular Biology book series (MIMB, volume 1186)


Extracellular stimuli activate, on target cells, a number of signal transduction processes regulating gene expression and the function and/or synthesis of the proteins. In order to highlight the slight changes of the quantity and quality of the proteome it is essential to optimize preparative strategies able to improve the signals of the less expressed proteins and to standardize the use of high-throughput techniques useful to detect them. We describe a complete workflow useful to enrich, from PBMC protein extracts and extrapolated to their subpopulations, the low-molecular-weight proteins and peptides and to detect them by SELDI-TOF protein profiling. The described protocol can also be applied to MALDI-TOF/MS instruments in order to obtain fast, reproducible, and high-quality protein profiles.

Key words



  1. 1.
    Lodish H, Berk A, Matsudaira P, Kaiser CA, Krieger M, Scott MP, Zipurksy SL, Darnell J (2004) Molecular cell biology, 5th edn. WH Freeman, New York, NYGoogle Scholar
  2. 2.
    Richards J, Le Naour F, Hanash S, Beretta L (2002) Integrated genomic and proteomic analysis of signaling pathways in dendritic cell differentiation and maturation. Ann N Y Acad Sci 975:91–100PubMedCrossRefGoogle Scholar
  3. 3.
    Hettinger J, Richards DM, Hansson J et al (2013) Origin of monocytes and macrophages in a committed progenitor. Nat Immunol 14(8):821–830PubMedCrossRefGoogle Scholar
  4. 4.
    Jeong HC, Lee SY, Lee EJ et al (2007) Proteomic analysis of peripheral T-lymphocytes in patients with asthma. Chest 132(2):489–496PubMedCrossRefGoogle Scholar
  5. 5.
    Papale M, Rocchetti MT, Gesualdo L (2013) Clinical proteomics: the potentiality of urine analysis for understanding diabetic nephropathy. EMJ Neph 1:32–39Google Scholar
  6. 6.
    Papale M, Di Paolo S, Magistroni R et al (2010) Urine proteome analysis may allow noinvasive differential diagnosis of diabetic nephropathy. Diabetes Care 33(11):2409–15PubMedCentralPubMedCrossRefGoogle Scholar
  7. 7.
    Sui W, Dai Y, Zhang Y et al (2010) Proteomic profiling of uremia in serum using magnetic bead-based sample fractionation and MALDI-TOF MS. Ren Fail 32(10):1153–9PubMedCrossRefGoogle Scholar

Copyright information

© Springer Science+Business Media New York 2014

Authors and Affiliations

  1. 1.Department of Surgery and Medical SciencesUniversity of FoggiaFoggiaItaly
  2. 2.Department of Emergency and Organ TransplantationUniversity of Bari Aldo MoroBariItaly

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