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 A 2 (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. Personalized medicine: Translation of personalized medicine from the laboratory into the clinic requires the development of new sensitive and quantitative methods for the measurements of clinically relevant markers. A new NMR method (R-ELISA) has been used to detect the folate receptor on human ovarian cancer cells (see picture) using a phospholipase that is able to induce the release of paramagnetic species from liposomes.
A Quantitative Relaxometric Version of the ELISA Test for the Measurement of Cell Surface Biomarkers
ALBERTI, DIEGO;STEFANIA, Rachele;RUGGIERO, MARIA ROSARIA;Marta Tapparo;GENINATTI CRICH, Simonetta;AIME, Silvio
2014-01-01
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 A 2 (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. Personalized medicine: Translation of personalized medicine from the laboratory into the clinic requires the development of new sensitive and quantitative methods for the measurements of clinically relevant markers. A new NMR method (R-ELISA) has been used to detect the folate receptor on human ovarian cancer cells (see picture) using a phospholipase that is able to induce the release of paramagnetic species from liposomes.
I documenti in IRIS sono protetti da copyright e tutti i diritti sono riservati, salvo diversa indicazione.
