Electrochemiluminescence immunosensor for cytokeratin fragment antigen 21-1 detection using electrochemically mediated atom transfer radical polymerization.

The cytokeratin fragment antigen 21-1 (CYFRA 21-1) protein is a critical tumor biomarker tightly related to non-small cell lung cancer (NSCLC). Herein, we prepared an effective electrochemiluminescence (ECL) immunosensor for CYFRA 21-1 detection using electrochemically mediated atom transfer radical polymerization (eATRP). The CYFRA 21-1 antigen was fixed on the electrode surface by constructing a sandwich type antibody-antigen-antibody immune system. The sensitivity of ECL was improved by using the eATRP reaction. In this method, eATRP was applied to CYFRA 21-1 detection antibody with N-acryloyloxysuccinimide as functional monomer. This is the first time that ECL and eATRP signal amplification technology had been combined. Under the optimized testing conditions, the immunosensor showed a good linear relation in the range from 1 fg mL-1 to 1 µg mL-1 at a limit of detection of 0.8 fg mL-1 (equivalent to ~ 134 molecules in a 10 µL sample). The ECL immunosensing system based on eATRP signal amplification technology provided a new way for rapid diagnosis of lung cancer by detecting CYFRA 21-1. The paper prepared an electrochemiluminescence biosensor for ultrasensitive detection of CYFRA 21-1 via eATRP signal amplification strategy, which had the advantages of high sensitivity, reproducibility, and held potential prospect for analysis of low-abundance.

Intramolecular photoinitiator induced atom transfer radical polymerization for electrochemical DNA detection.

A novel electrochemical biosensor was reported for the first time to achieve highly sensitive DNA detection based on photoinduced atom transfer radical polymerization (photoATRP). In this work, PNA was applied as the capture probe to specifically recognize the target DNA (TDNA), and we utilized lung cancer DNA as TDNA. The ATRP initiator was introduced to the electrode surface via phosphate-Zr4+-carboxylate chemistry. PhotoATRP was activated under blue light irradiation based on a photoinitiator I2959, which produced free radicals via homolytic cleavage. Subsequently, Cu2+ was reduced to Cu+ with the assistance of the free radicals, and numerous electroactive probes were grafted onto the electrode surface. Under optimal conditions, the limit of detection (LOD) of this method was 3.16 fM (S/N = 3, R2 = 0.992), and the linear range was from 10 fM to 1.0 nM. More importantly, the preparation process of this biosensor was simple and less laborious with a low background signal, suggesting good potential in practical applications.

Dual signal amplification by polysaccharide and eATRP for ultrasensitive detection of CYFRA 21-1 DNA.

Cytokeratin fragment antigen 21-1 (CYFRA 21-1) DNA is a crucial biomarker closely associated with non-small cell lung cancer. Here, we fabricated a novel electrochemical biosensor for ultrasensitive detection of CYFRA 21-1 DNA via polysaccharide and electrochemically mediated atom transfer radical polymerization (eATRP) dual signal amplification. Specifically, thiolated peptide nucleic acid (PNA) probes at 5'-terminals are immobilized on the gold electrode surface for specific recognition of CYFRA 21-1 DNA (tDNA). After hybridization, hyaluronic acid (HA) is linked to the hybridized PNA/DNA duplexes via the recognized carboxylate-Zr4+-phosphate chemistry. Then multiple initiators of the polymerization reaction are introduced via esterification reaction. Lastly, large numbers of electro-active monomers are successfully grafted from the initiation sites of functionalized HA by eATRP reaction, significantly amplifying the electrochemical signal. Under optimal conditions, the constructed sensor can detect as low as 9.04 aM tDNA. Further, this proposed biosensor can also be applied to the direct detection of double-stranded DNA (dsDNA), obtaining 0.12 fM as the detection limit. Besides, this strategy shows high selectivity for mismatched bases and excellent applicability for CYFRA 21-1 DNA detection in the serum samples. Given its high sensitivity, selectivity, ease of operation, low cost and environmental friendliness, this biosensor has considerable potential in early diagnosis and biomedical application.

Metal-Free Photoinduced Atom Transfer Radical Polymerization for Highly Sensitive Detection of Lung Cancer DNA.

Convenient and sensitive detection of biomolecules is of great significance to disease diagnosis. In this work, a metal-free photoinduced atom transfer radical polymerization (photoATRP) by a reductive quenching pathway as a novel strategy is applied to achieve lung cancer DNA detection. Thiolated PNA is exploited to specifically recognize target DNA, and the initiator of photoATRP is linked to the electrode surface via phosphate-Zr4+ -carboxylate. Under the excitation of blue light, the reductive quenching pathway is activated with eosin Y (EY) as photoredox catalyst and N,N,N',N'',N'-pentamethyldiethylenetriamine (PMDETA) as electron donor, and numerous polymeric chains are formed. Under optimal conditions, the linear range of this strategy is from 0.1 pm to 10 nm (R2 =0.989) with a limit of detection (LOD) of 1.4 fm (14 zmol in 10 µL). The variety of possible light sources for photoATRP and simple operation endow this biosensor with great potential for practical applications.