A Quantitative Relaxometric Version of the ELISA Test for the Measurement of Cell Surface Biomarkers |
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| Authors: | Dr Diego Alberti Dr Mark van't?Erve Dr Rachele Stefania Dr Maria Rosaria Ruggiero Dr Marta Tapparo Dr Simonetta Geninatti?Crich Prof Silvio Aime |
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| Institution: | Department Molecular Biotechnology and Health Sciences, Institution University of Torino, Address via Nizza 52, Torino (Italy) |
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| Abstract: | Quantitative measurement of marker expression in diseased cells is still a topic of considerable interest and different methodologies are currently under intense scrutiny. This work aims at developing an in vitro diagnostic method based on the release of paramagnetic species from relaxometrically “silent” liposomes operated by the action of a phospholipase A2 (PLA2) previously targeted to the epitope of interest. The released paramagnetic species causes an increase of the longitudinal water proton relaxation rate proportional to the number of PLA2 bound to the cell outer surface. The sensitivity of the herein proposed method, named R‐ELISA, was attempted in the detection of folate receptor expression on human ovarian cancer cells by functionalizing PLA2 with folic acid. Receptor/cell number of 8.3×105 has been measured on IGROV‐1 cells. The R‐ELISA assay can detect nanomolar cell suspension receptor concentrations and has been validated by well‐established spectrofluorimetric procedures. |
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| Keywords: | analytical methods NMR spectroscopy paramagnetic liposomes personalized medicine relaxometry |
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