Abstract
In this work, we have developed an assay for the detection of proteins by functionalized nanomaterials coupled with laser-induced desorption/ionization mass spectrometry (LDI-MS) by monitoring the generation of metal cluster ions. We achieved selective detection of three proteins [thrombin, vascular endothelial growth factor-A165 (VEGF-A165), and platelet-derived growth factor-BB (PDGF-BB)] by modifying nanoparticles (NPs) of three different metals (Au, Ag, and Pt) with the corresponding aptamer or antibody in one assay. The Au, Ag, and Pt acted as metal bio-codes for the analysis of thrombin, VEGF-A165, and PDGF-BB, respectively, and a microporous cellulose acetate membrane (CAM) served as a medium for an in situ separation of target protein-bound and -unbound NPs. The functionalized metal nanoparticles bound to their specific proteins were subjected to LDI-MS on the CAM. The functional nanoparticles/CAM system can function as a signal transducer and amplifier by transforming the protein concentration in...
Highlights
The greatest challenge in the detection of specific proteins or tumor markers for the diagnosis of cancer is their low concentrations in human plasma.[1,2] Interferences due to other proteins with similar properties cause difficulties in their selective detection by conventional methods.[2]
To resolve the interference from background proteins and the unpredictable fragmentation of analytes encountered in SALDI-mass spectrometry (MS), we have developed a simple immunoassay that exhibits significant potential for the simultaneous detection of different proteins in a single analysis by monitoring the cluster ion signals generated from the metal nanoparticles (NPs) under pulse laser irradiation
We used cellulose acetate membrane (CAM), which can be directly mounted onto a plate for LDI-MS analysis, to serve as a medium to separate the antibody (Ab)- or aptamer (Apt)-modified NPs and their conjugates formed with their targeting proteins in situ
Summary
The greatest challenge in the detection of specific proteins or tumor markers for the diagnosis of cancer is their low concentrations in human plasma.[1,2] Interferences due to other proteins with similar properties cause difficulties in their selective detection by conventional methods.[2]. The functionalization of different metal NPs with their respective aptamers or antibodies enables specific targeting of thrombin, VEGF-A165, and PDGF-BB (Fig. 1).
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