In this paper, a simple, ultrasensitive and label-free electrochemical α2,6-sialylated glycans biosensor based on reduced graphene oxide-tetraethylene pentamine-1-butyl-3-methylimidazolium hexafluorophosphate (BMIMPF 6 ) hybrids was developed. Due to the abundance of amino groups from reduced graphene oxide-tetraethylene pentamine (rGO-TEPA) and the electrostatic interaction of BMIMPF 6 , bimetallic gold platinum alloy nanoparticles (AuPtNPs) were densely adsorbed onto the surface of the nanocomposite , providing a large surface area available for the immobilization of Sambucus nigra agglutinin (SNA). AuPtNPs have excellent conductivity and catalytic activity , which can promote electron transfer between the electrolyte solution and the surface of electrode and can enhance the sensitivity of biosensor. SNA, which specifically binds α2,6-sialylated glycans, was covalently immobilized on AuPtNPs for specific detection of α2,6-sialylated glycans in human serum. Under optimal experimental conditions, amperometric response changes were used to detect α2,6-sialylated glycans with a broad linear range of 10 fg mL −1 –1 μg mL − 1 and a low detection limit of 3 fg mL − 1 ( S / N =3). When applied to spiked serum samples, the recovery of the developed biosensor ranged from 100.8% to 101.4%, suggesting that the electrochemical biosensor would be suitable for the practical detection of α2,6-sialylated glycans.
α2,6-Sialylated glycans are crucial molecular targets for cancer diagnosis and clinical research. In this work, a novel ultrasensitive electrochemical biosensor was fabricated based on a graphite oxide (GO), Prussian blue (PB), and PTC-NH 2 (an ammonolysis product of 3,4,9,10-perylenetetracarboxylic dianhydride) nanocomposite for the selective detection of α2,6-sialylated glycans. To increase the sensitivity of the electrochemical biosensor, gold nanoparticles (GNPs) were immobilized on a GO–PB–PTC-NH 2 modified glassy carbon electrode (GCE). Sambucus nigra agglutinins (SNAs), which specifically bind with α2,6-sialylated glycans, were covalently immobilized on GNPs for the sensitive detection of α2,6-sialylated glycans in serum. This proposed method can be applied to human serum, and it worked well over a broad linear range (0.1 pg mL −1 –500 ng mL −1 ) with detection limits of 0.03 pg mL −1 . Moreover, recovery of the spiked samples ranged from 100.2% to 105.0%, suggesting that this excellent electrochemical biosensor can be used for the practical detection of α2,6-sialylated glycans.
Sensitive detection of tumor biomarkers is crucial for early diagnosis and prognosis evaluation of cancer. Owing to no need of labelled antibody, formation of sandwich immunocomplexes and additional solution-based probe, probe-integrated electrochemical immunosensor for reagentless detection of tumor biomarkers is highly desirable. In this work, sensitive and reagentless detection of a tumor biomarker is realized based on fabrication of a probe-integrated immunosensor by confining redox probe in electrostatic nanocage array modified electrode. Indium tin oxide (ITO) electrode is employed as the supporting electrode because it is cheap and easily available. The silica nanochannel array consisted of two layers with opposite charges or different pore diameters was designated as bipolar films (bp-SNA). In this work, Electrostatic nanocage array is equipped on ITO electrode by growth of bp-SNA with two layered nanochannel array having different charge properties including a negatively charged silica nanochannel array (n-SNA) and a positively charged amino-modified SNA (p-SNA). Each SNA can be easily grown with 15 s using electrochemical assisted self-assembly method (EASA). Methylene blue (MB) is applied as the model electrochemical probe with positive charge to be confined in electrostatic nanocage array with stirring. The combination of the electrostatic attraction from n-SNA and the electrostatic repulsion from p-SNA endows MB with highly stable electrochemical signal during continuous scanning. When the amino groups of p-SNA are modified using the bifunctional glutaraldehyde (GA) to introduce aldehydes, the recognitive antibody (Ab) of the most commonly used tumor biomarker, carcinoembryonic antigen (CEA), can be covalently immobilized. After the non-specific sites are blocked, the immunosensor is successfully fabricated. As the formation of antigen-antibody complex decreases electrochemical signal, the immunosensor can achieve reagentless detection of CEA ranged from 10 pg/mL to 100 ng/mL with a low limit of detection (LOD, 4 pg/mL). Determination of CEA in human serum samples is realized with high accuracy.
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