In the present work we design a novel aptamer-based silver nanosensor for one-spot simultaneously detection of multiple proteins. SS-DNA modified AgNPs were immobilized on the aldehyde coated glass slide to form an AgNP array. Then dye-labeled aptamer sequences were allowed to hybridize with their complementary strands assembled on the surface of AgNPs. The target proteins were introduced to associate with the corresponding aptamers to form the aptamer–target complexes. The removal of the aptamer–target complexes resulted in a remarkable decrease in fluorescent signals. This nanosensor is found to be highly sensitive for the detection of proteins. When thrombin was employed as a sample model, the limit of detection of the optimized nanosenor was 0.4 fmol with a linear response of 0.8 fmol to 0.5 pmol. We further demonstrated the multiple protein detection of IgE and thrombin using multicolor silver nanoprobes, which shows effective recognition of the relative protein individually or simultaneously. This silver nanosensor offers a unique heterogeneous approach for protein detection with several advantages, such as high sensitivity, rapidity, high throughput, and miniaturization.
