Erythropoietin: We are currently working on some endogenous substances that are recombinantly produced by the pharmaceutical industry, particularly erythropoietin (EPO) (INN: epoetin) and NESP (INN: darbepoetin) with the view of characterising their differences with respect to the endogenously produced hormone. In particular we are interested in studying differences in glycosylation, observed by isoelectric focusing but still not explained. The reports of autoimmune problems caused by the administration of those pharmaceutical preparations shows that the glycosylation profiles, main difference between the exogenous and endogenous product may be critical from the safety point of view. This characterisation implies the development of methodologies also applicable to other fields of biochemical analysis or other substances under similar circumstances.
We are also currently trying to develop monoclonal antibodies against N-glycolyl-neuraminic acid (Neu5GC), a monosaccharide only present in the recombinant material with the intention of developing a sensitive technique (ELISA, immuno PCR, RT-immuno PCR).
Future plans are based on the developments of instrumental (basically MS) methodologies to detect the presence of modifications on the glycosylation of proteins (e.g. sulfation, phosphorylation) most probably the moieties responsible for the differences between endogenous and exogenous EPO. The particular urinary excretion of EPO and how those changes are introduced following its secretion from the kidneys will also be object of our interest.
Gene Therapy is advancing as one of the important therapies in the XXI century. To detect a correct application of gene therapy and consequently the expression of the target protein is of paramount importance for the success of the approach. A non-invasive detection method for successful gene therapy application would be highly desirable. In sport, the diagnostic detection of a gene therapy application could also be used to deter the malpractice of such an important medical tool (gene doping). The project Imagene is addressed to gain insight into such a diagnostic capability, in coordination with leading groups in genetics and imaging.
Regarding blood misuse, the objective is to develop methods to detect the abuse of blood transfusions in sportsmen. Two approaches will be studied. The first one is based on the detection of contaminants present in bags used to store blood or red blood cell concentrates; the concentrations of these contaminants and their metabolites should be higher in body fluids of individuals subjected to transfusion compared to non-transfused subjects. The second approach is based on the detection of markers of red blood cell aging due to storage of blood in transfused subjects.