Pulsed-Laser Desorption/Ionization of Clusters from Biofunctional Gold Nanoparticles: Implications for Protein Detections
journal contributionposted on 24.10.2012, 00:00 by Yin-Chun Liu, Huan-Tsung Chang, Cheng-Kang Chiang, Chih-Ching Huang
In this paper, we describe a pulsed-laser desorption/ionization mass spectrometry (LDI-MS) approach for the detection of proteins with femtomolar sensitivity through the analysis of gold (Au) clusters desorbed from aptamer-modified gold nanoparticles (Apt–AuNPs) on a nitrocellulose membrane (NCM). After the target protein (thrombin) was selectively captured by the surface-bound 29-mer thrombin-binding aptamer (TBA29), the thrombin/TBA29–AuNP complexes were concentrated and deposited onto the NCM to form a highly efficient background-free surface-assisted LDI substrate. Under pulsed laser irradiation (355 nm), the binding of thrombin decreased the desorption and/or ionization efficiencies of the Au atoms from the AuNP surfaces. The resulting decreases in the intensities of the signals for Au clusters in the mass spectra provided a highly amplified target-labeling indicator for the targeted protein. Under optimized conditions, this probe was highly sensitive (limit of detection: ca. 50 fM) and selective (by at least 1000-fold over other proteins) toward thrombin; it also improved reproducibility (<5%) of ion production by presenting a more-homogeneous substrate surface, thereby enabling LDI-based measurements for the accurate and precise quantification of thrombin in human serum. This novel LDI-MS approach allows high-speed analyses of low-abundance thrombin with ultrahigh sensitivity; decorating the AuNP surfaces with other aptamers also allowed amplification of other biological signals.