Microbubble Enzyme-Linked ImmunoSorbent Assay (MB-ELISA)

Project Description

Ultrasound (US) microbubbles (MB) can be targeted to specific proteins present on the tumor endothelium enabling for US molecular imaging. The development of an US molecular imaging protocol is highly dependent on the characteristics of the employed targeted contrast agents. Together with the selection and validation of the molecular target, the preparation and in vitro characterization of the contrast agent are highly critical steps for successful translation into in vivo studies and therefore require robust protocols for the assessment of MB affinity, sensitivity and specificity. In vitro validation approaches from different laboratories are very diverse and rely either on partial microscopic analysis or on complex in vitro US imaging setups. We propose an enzymatic detection approach that allows the assessment of MB binding specificity in a reliable, reproducible, robust, high-throughput-like, and quantitative manner (Figure 1).

The developed MB-ELISA provides a format for comparison of a high number of samples in a rapid protocol which is required for contrast agents like MB which are unstable in time. In addition to the quick enzymatic detection and revelation protocol, the assay does not need any time-consuming image analysis providing results in a shorter time delay than with microscopy analysis, which is today the gold standard for analysis of attached MB in in vitro assays. Microscopy further showed that MB distributed heterogeneously over the surface of the wells underlining the issue of the selection of representative fields of view and the need for whole-well analysis. So far, enzymatic detection is the first technique addressing this issue.

Figure 1: Working principle of MB-ELISA.


LabTAU: Jennifer Wischhusen (PhD student), Dr. Frederic Padilla (CNRS researcher)