SELDI Proteomics Approach to Identify Proteins Associated with T-Cell Clone Senescence

  • Dawn J. Mazzatti
  • Robin Longdin
  • Graham Pawelec
  • Jonathan R. Powell
  • Rosalyn J. Forsey
Part of the Medical Intelligence Unit book series (MIUN)


The immune system undergoes many complex changes as a result of the aging process. Elderly humans have altered cellular redox levels and deregulated immune responses, both key events underlying the progression of chronic degenerative diseases of aging, such as atherosclerosis and Alzheimer’s disease. T-cells are one of the major cell types affected by aging. As such, indentifying bio-markers of T-cell aging and senescence would aid in identifying and developing novel intervention strategies.

Proteomics has emerged recently as a rapidly expanding and innovative field, investigating protein expression, interactions, localisation and function at a global level. In this context, we used the Ciphergen Protein Chip PCS4000 surface enhanced laser desorption/ionisation (SELDI) system, a combination of affinity chromatography and mass spectrometry, to study protein profile changes that occur during in vitro T-cell aging and immunosenescence. This technology offers faster, higher throughput analysis of protein expression than the more traditional 2-dimensional-gel electrophoresis method, allowing the screening of larger sample numbers for potential bio-markers. Furthermore, on-chip affinity chromatography reduces sample complexity and permits targeting of protein groups.

Ciphergen H50 chips (reversed-phase chromatography) and Q10 chips (anion-exchange chromatography) were used to target hydrophobic and negatively charged proteins, respectively, in T-cell lysates. Biomarker analysis using the CiphergenExpress software identified differential expression of a variety of peaks associated with in vitro T-cell aging. A consistent pattern of differential protein expression was observed between both early and late passage T-cell clones grown in vitro and from T-cell clones derived from young and old donors. The corresponding proteins were identified by a combination of SELDI-TOF-MS, peptide mass fingerprinting MALDI-TOF-MS and Nanospray-IonTrap-MS/MS. Various molecules were demonstrated to be differentially expressed in aging and senescence, with good correlation between SELDI and MALDI data. This suggests that SELDI is a valuable tool in elucidating proteomic differences between cell populations, identifying potential biomarkers in large sample populations that can then be investigated further using more sensitive MS/MS methods for identification.


Peptide Mass Fingerprinting Late Passage HPLC Fraction Young Donor MOWSE Score 
These keywords were added by machine and not by the authors. This process is experimental and the keywords may be updated as the learning algorithm improves.


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Copyright information

© Landes Bioscience and Springer Science+Business Media 2007

Authors and Affiliations

  • Dawn J. Mazzatti
    • 3
  • Robin Longdin
    • 1
  • Graham Pawelec
  • Jonathan R. Powell
    • 2
  • Rosalyn J. Forsey
    • 1
  1. 1.LCG BioscienceCambridgeUK
  2. 2.Unilever Corporate ResearchSharnbrookUK
  3. 3.Unilever Corporate ResearchSharnbrookUK

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