Radiation-functionalized polyolephinic films for the isolation of rare cells in biological specimens for diagnostic applications

Abstract

Early detection and effective treatment significantly impact survival rates in disease management. In prenatal diagnostics, micromanipulation is a dependable method for manually isolating rare fetal cells from maternal fluids for molecular or cytogenetic analysis. While effective, this approach is costly and time-consuming due to the need for skilled personnel and specialized equipment1. This study aims to enhance the efficiency and affordability of prenatal diagnosis in hospital settings by developing a device for semi-automated selection of rare cells from biological samples. The device utilizes polyolephinic films functionalized to obtain carboxyl groups on the surface via electron-beam radiation. These functional groups can be then used to decorate the films with antibodies to selectively capture target cells based on surface antigens. Bioconjugation methods employing EDC/NHS chemistry were developed to impart cell-capture capabilities to the films, exploiting the streptavidin or avidin with biotin lock and key bound. Fluorescently labelled antibodies enabled assessment of conjugation success via fluorimetry and spectrofluorimetry. Cell capture tests were conducted using mesenchymal stem cells (MSC) on antibody-decorated films, with observations made using optical and confocal microscopy to evaluate capture efficacy. Release after capture mechanism of the streptavidin or avidin with biotin lock and key bound was also tested using fluorescent biotinylated probes.