Protein detection in blood with single-molecule imaging

The ability to characterize individual biomarker protein molecules in patient blood samples could enable diagnosis of diseases at an earlier stage, when treatment is typically more effective. Single-molecule imaging offers a promising approach to accomplish this goal. However, thus far, single-molecule imaging methods have not been translated into the clinical setting. The detection limit of these methods has been confined to the picomolar (10-12 M) range, several orders of magnitude higher than the circulating concentrations of biomarker proteins present in many diseases. Here, we describe single-molecule augmented capture (SMAC), a single-molecule imaging technique to quantify and characterize individual protein molecules of interest down to the subfemtomolar (<10(-15) M) range. We demonstrate SMAC in a variety of applications with human blood samples, including the analysis of disease-associated secreted proteins, membrane proteins, and rare intracellular proteins. SMAC opens the door to the application of single-molecule imaging in noninvasive disease profiling.

Automated summary

Characterizing individual biomarker protein molecules in patient blood samples using single-molecule imaging techniques holds promise for early disease diagnosis and effective treatment. However, current methods have detection limits that are orders of magnitude higher than needed, making them unsuitable for clinical use. The development of SMAC, with its ability to quantify and characterize protein molecules down to the subfemtomolar range, presents a new avenue for noninvasive disease profiling with single-molecule imaging.