The Construction of Protein Biochip Arrays Using Surface Protein Resonance Optical Detection

Specific goal: to evaluate the sensitivity and specificity of the molecular sensing tools developed in the project (SPR and SERS) for the observation and quantification of individual proteins and protein complexes related to onco-hematological diseases and to identify disease-associated mechanisms by validating microRNA and mRNA profiling data of patients and healthy individuals at a protein level using SPR biosensors.

Cellular functions are the result of the coordinated action of groups of proteins interacting in molecular assemblies or pathways. The formation of blood cellular components, hematopoiesis, is regulated at DNA, RNA and protein levels by balanced processes that, if dislocated, may give rise to a blood tumor. Studies of multiprotein complex formation are likely to play a major role in determining cellular abnormalities in onco-hematological diseases, such as chronic myeloid leukemia.

The crucial mechanisms that still remain to be discovered are those causing hematologic tumors, which are exceptionally aggressive, refractory to therapy or frequently relapsing. The elucidation of protein–protein interaction relationships is an essential step towards the understanding of the complex processes behind the tumor onset and development. In addition, there is an ongoing search for specific plasmatic biomarkers the levels of which could be correlated with specific disorders. This is a tremendously challenging task also due to the heterogeneity of hematological malignancies.

SPR biosensors provide a powerful tool for real-time monitoring of protein–protein interactions and therefore have been used to investigate various aspects of onco-hematological diseases, such as complex processes in cells, interactions of proteins including grow factors, inhibitors of cell progression, cytokines and other proteins. The SPR has also been demonstrated to have great potential for the quantification of protein biomarkers of hematologic tumors.

The SERS method has also been used for protein detection, providing information about the protein structure and its changes upon interaction with other proteins and therapeutics. SERS has been used in several structural studies of proteins related to acute myeloid leukemia and involved in chromatin modifications, or of proteins with tumor suppressor function. The potential of SERS as a diagnostic method has been demonstrated in vivo and in vitro for example in the detection of circulating tumor cells in blood, B-cell lymphoma 2 gene, and the detection of hematologic malignancy biomarkers.