Identification of new diagnostic markers

Diagnosis and disease monitoring are crucial aspects of medicine. Biomarkers are   important for diagnosis and the clinical follow-up of patients, and because their use is relatively cheap and non-invasive, they hold great interest for the scientific community.

Biomarkers can be identified through a number of approaches:

  • genetic diagnostic kits for rare and multifactorial diseases based on multiplex PCR techniques that can identify mutations that are relatively common in the general population;
  • genotyping to identify rare or frequent variants using novel high-throughput deep sequencing techniques;
  • transcriptomic approaches that assess the expression profile of specific diseases using microarrays and deep sequencing in order to identify altered genes that might be suitable as biomarkers;
  • epigenomic approaches that map DNA methylation and histone modifications genome-wide;
  • proteomic approaches that identify protein biomarkers in tissues and biological fluids through DIGE associated with the MS peptide fingerprinting.

Protein chip associated with Surface Enhanced Laser Desorption Ionization Time of Flight (SELDI-TOF) is a unique method that uses chromatographic surfaces (or chips) to bond proteins and peptides on the basis of their physical and chemical properties; the chips are then analyzed with a series 4000 di PBSIIc TOF-MS reader (Ciphergen Biosystems, Inc). The extracted proteins are separated on arrays of diverse binding surfaces made up of different hydrophobic components (H5, H50), cation/anion exchangers (Q10, CM10), affinities for metal ions (IMAC), etc. Surfaces can be also made to specifically bind to antibodies, receptors and ligands (PS10 and PS20). After a wash, the bound proteins are analyzed with a total spectrometer (MS).

This technique is highly sensitive and is therefore ideal for the analysis of small sample volumes and low-molecular-weight molecules. It also allows semiquantitative and statistical analyses, and can help in the setting up of purification steps prior to fingerprinting or mass sequencing.

With this technique, BioTekNet is able to:

  • identify markers for the diagnosis and classification of neoplasms, such as prostate cancer;
  • identify (within tissues and biological fluids) and validate genetic (polymorphic), proteic and transcriptomic risk factors for infarction, ictus, hypertension, etc.;
  • identify molecular and genetic markers of cardiovascular diseases and alterations in correlated quantitative parameters through the analysis of genealogical, clinical and genetically characterised isolated populations.

 

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