Ferritin-Based Electrochemiluminescence Nanosurface Energy Transfer System for Procalcitonin Detection Using HWRGWVC Heptapeptide for Site-Oriented Antibody Immobilization

Abstract

In this study, an ultrasensitive biosensor based on a novel electrochemiluminescence nanosurface energy transfer (ECL-NSET) system was developed for procalcitonin (PCT) detection. N-(Aminobutyl)-N-(ethylisoluminol) (ABEI) as ECL donor was externally crosslinked with ferritin (ABEI-Ft), whose ECL emission was boosted by a ferritin nanocore in the presence of hydrogen peroxide. Then single gold nanoparticles (Au NPs) as ECL acceptor were in situ reduced on the ABEI-Ft surface to fabricate a donor−acceptor nanostructure (ABEI-Ft@ Au). ECL quenching that occurred in ABEI-Ft@Au correlates well with the NSET theory rather than Forster resonance energy transfer (FRET). To improve sensitivity of the biosensor, HWRGWVC heptapeptide (HWR) was utilized to capture the antibody Fc portion via specific interaction to realize site-oriented immobilization. After connecting with ABEI-Ft@Au via Au−S bonding, HWR improved the incubation efficiency of the antibody with a better maintained biological activity. Under optimal conditions, the proposed biosensor provided a quantitative readout to PCT concentration in the range of 100 fg/mL to 50 ng/mL with a detection limit of 41 fg/mL. With favorable specificity, stability, and reproducibility, this highly efficient ECL-NSET system will inspire more interest in nanometal-based ECL quenching mechanism studies and development of new methods for site-oriented antibody immobilization to improve sensitivity.

Integrating Highly Efficient Recognition and Signal Transition of g-C3N4 Embellished Ti3C2 MXene Hybrid Nanosheets for Electrogenerated Chemiluminescence Analysis of Protein Kinase Activity

“…The high affinity of antibody–antigen improves the selectivity of biosensors. , However, these immunoassays generally suffered from false positive analysis, poor storage stability, and cross-link reaction from the intrinsic immunoreactions. Recently, electrogenerated chemiluminescence has received much attention, which is a light emission process in a redox reaction of electrogenerated reactants and combines the advantages of the electrochemistry and luminescence with high sensitivity, simple instruments, easy miniaturization, and low cost . Moreover, the electrogenerated chemiluminescence (ECL) technique is a potential method and is spatial controlling, and has potential for the high-throughput analysis .…”

Triple Amplification of 3,4,9,10-Perylenetetracarboxylic Acid by Co2+-Based Metal–Organic Frameworks and Silver–Cysteine and Its Potential Application for Ultrasensitive Assay of Procalcitonin

“…Meanwhile, the AgCys complex was synthesized to catalyze S 2 O 8 2– to produce more SO 4 •‑ , thus amplifying the ECL signal once again. Furthermore, HWRGWVC (HGC), a heptapeptide with many advantages, such as low cost, simple preparation, and excellent stability, was introduced into the biosensor, which could combine the Fc fragments of an antibody by Au–S bonding to achieve the fixed point fixation . Meanwhile, the incubation efficiency of the antibody was improved with the introduction of HGC, which enhanced the sensitivity of the biosensor.…”

“…Furthermore, HWRGWVC (HGC), a heptapeptide with many advantages, such as low cost, simple preparation, and excellent stability, was introduced into the biosensor, which could combine the Fc fragments of an antibody by Au−S bonding to achieve the fixed point fixation. 20 Meanwhile, the incubation efficiency of the antibody was improved with the introduction of HGC, which enhanced the sensitivity of the biosensor. With superior sensitivity and bioactivity, the proposed triple-amplified biosensor realized an ultrasensitive assay of PCT in human serum.…”

Advancement in biosensors for inflammatory biomarkers of SARS-CoV-2 during 2019–2020

“…Yang et al fabricated an electrochemiluminescence sensor for PCT ( Yang et al, 2019 ). In this work, N-(Aminobutyl)-N-(ethylisoluminol) (ABEI) linked with ferritin acted as a donor molecule, whereas the single gold nanoparticle acted as the acceptor molecule.…”

“… 3.3–125 μg/mL 1.95 μg/mL Giorgi-Coll et al (2019) Optical C.I. 1.25–9000 ng/mL 0.38 ng/mL Sene Ingridi de Souza et al (2020) Optical Reflectance 50–350 pg/mL 0.52 and 0.08 pg/mL Khan et al (2020) Procalcitonin Electro/optical EL 100 fg/mL to 50 ng/mL 41 fg/mL Yang et al (2019) Electrochemical Amperometry 0.01 pg/mL to 10 ng/mL 0.002 pg/mL Liu et al (2019) Electrochemical Impedimetry 0.01–10 ng/mL 0.10 ng/mL Tanak et al (2019) Electrochemical DPV 0.001–25 ng/mL 3 × 10 −4 ng/mL Gao et al (2020) Electrochemical Amperometry 10 pg/mL to 50 ng/mL 0.15 pg/mL Abbas and Soomro (2019) Electrochemical DPV 0.01–1.0 pg/mL 2 fg/mL Medetalibeyoglu et al (2020) Electrochemical Amperometry 500 fg/mL to 50 ng/mL 130 fg/mL Qu et al (2020) Electrochemical ...…”