A Quantitative Relaxometric Version of the ELISA Test for the Measurement of Cell Surface Biomarkers

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 invitro diagnostic method based on the release of paramagnetic species from relaxometrically silent liposomes operated by the action of a phospholipase A(2) (PLA(2)) 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 PLA(2) 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 PLA(2) with folic acid. Receptor/cell number of 8.3x10(5) 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.