Currently,
it remains challenging to count protein-biomarker molecules
present in a small droplet of biological samples. Herein, we propose
a gold nanoparticle (GNP) probe-assisted sandwich-counting strategy
that relies on a GNP probe, an antibody-functionalized chip to “count”
antigen molecules using a scanning electron microscope. Both standard
carcinoembryonic antigen (CEA) and two real CEA-related tumor samples
(tumor tissues and serum) were assayed to demonstrate the proof-of-concept
of the counting strategy. Results show that our method is excellently
correlative with enzyme-linked immuno-sorbent assay (ELISA) that is
widely used in clinics for antigen or antibody detection and the limit
of detection of our enumeration strategy reaches down to 0.045 ng/mL,
which is ∼40 times more sensitive than the conventional ELISA.
Therefore, our GNP probe-assisted sandwich-counting strategy has the
potential to be used for quantification of protein biomarkers at ultralow
concentrations in early tumor specimens and detection of target proteins
in much diluted concentrations.