Bimodal Multiplexed Detection of Tumor Markers in Non-Small Cell Lung Cancer with Polymer Dot-Based Immunoassay

Abstract

Semiconducting polymer nanoparticles (Pdots) have been demonstrated to be a promising class of probes for use in fluorometric immunochromatographic test strips (ICTS). The advantages of Pdots in ICTSs include ultrahigh brightness, minimal nonspecific adsorption, and multicolor availability, which together contribute to the high sensitivity, good specificity, and multiplexing ability. These unique properties can therefore circumvent several significant challenges of commercial ICTSs, including insufficient specificity/sensitivity and difficulty in quantitative and multiplexed detection. Here, we developed a colorimetric and fluorescent bimodal readout ICTS based on gold-Pdot nanohybrids for the determination of carcinoembryonic antigen (CEA) and cytokeratin 19 fragment (CYFRA 21-1) expressed abnormally in human blood of non-small-cell lung cancer (NSCLS). The vivid color from Au nanomaterials can be used for rapid qualitative screening (colorimetry) in 15 min, while the bright fluorescence of Pdots is ideal for the advanced quantitative measurements of CEA and CYFRA21-1 concentrations in whole blood samples. This bimodal ICTS platform possesses phenomenal detection sensitivity of 0.07 and 0.12 ng/mL for CYFRA21-1 and CEA, respectively. The accuracy and reliability of this ICTS platform were further evaluated with clinical serum samples from NSCLS patients at different stages, showing good consistency with the results from electrochemiluminescence immunoassay.

On-bead DNA synthesis triggered by allosteric probe for detection of carcinoembryonic antigen

“…We used carcinoembryonic antigen (CEA), a biomarker of multiple cancers [ 13 , 20 – 22 ], as a protein model for testing. The DNA sequence of the recognition sequence (5′ATACCAGCTTATTCAATT 3′) in AP was reported by Tabar, H. et al [ 23 ].…”

“…For instance, enzyme-linked immunosorbent assay (ELISA), as a gold standard method for protein quantification, can hardly detect proteins of lower than 1 ng/mL [ 12 ]. Recently developed techniques, such as electrochemical assays [ 13 , 14 ] and Surface-enhanced Raman scattering [ 15 ], can lower the detection limit to 0.01–0.1 ng/mL. Yet, the poor repeatability and quantifiability of the methods have confined themselves to the laboratory.…”

Semiconducting polymer dots as fluorescent probes forin vitrobiosensing

“…Over the past few years, Yang-Hsiang Chan and his coworkers have employed Pdots in immunochromatographic test strips (ICTS). 69–72 Compared with conventional signal reporting reagents, such as colloidal gold nanoparticles, there are several advantages for Pdot-based ICTS: relatively high fluorescence brightness, high sensitivity and selectivity, excellent photostability, and multicolor emissions based on the efficient donor–acceptor system and facile surface functionalization.…”

“…Recently, Yang et al employed Au@Pdot-Based ICTS in the simultaneous detection of Carcinoembryonic antigen (CEA) and cytokeratin 19 fragment (CYFRA21-1) in human whole blood. 72 In their work, they prepared two Au@Pdot nanohybrids to detect CEA and CYFRA21-1, respectively, on a single test line. In particular, they coated Au nanorods with PFCN Pdots while they coated Au nanoparticles with PFTC6FQ Pdots.…”

Magnetic-Fluorescent Nanocluster Lateral Flow Assay for Rotavirus Detection

“…The development of POCT biosensors, specifically LFA, has received significant attention because of its rapid, simple, and inexpensive detection method. , However, commercial LFA kits have poor sensitivities and specificities compared to other immunoassay methods, such as RT-PCR or ELISA. An efficient way to increase the virus detection sensitivity is by integrating nanotechnologies with LFA diagnostics, including gold nanoparticles (NPs), quantum dots (QDs), upconversion NPs, and polymer dot-based LFA . Developing a rapid diagnostic toolkit with high sensitivity and specificity is thus pivotal to achieving more precise and accurate point-of-care diagnostics for a healthier, safer world …”