Ultrasensitive Electrochemical Detection of Glycoprotein Based on Boronate Affinity Sandwich Assay and Signal Amplification with Functionalized SiO₂@Au Nanocomposites

Abstract

Herein we propose a multiple signal amplification strategy designed for ultrasensitive electrochemical detection of glycoproteins. This approach introduces a new type of boronate-affinity sandwich assay (BASA), which was fabricated by using gold nanoparticles combined with reduced graphene oxide (AuNPs-GO) to modify sensing surface for accelerating electron transfer, the composite of molecularly imprinted polymer (MIP) including 4-vinylphenylboronic acid (VPBA) for specific capturing glycoproteins, and SiO nanoparticles carried gold nanoparticles (SiO@Au) labeled with 6-ferrocenylhexanethiol (FcHT) and 4-mercaptophenylboronic acid (MPBA) (SiO@Au/FcHT/MPBA) as tracing tag for binding glycoprotein and generating electrochemical signal. As a sandwich-type sensing, the SiO@Au/FcHT/MPBA was captured by glycoprotein on the surface of imprinting film for further electrochemical detection in 0.1 M PBS (pH 7.4). Using horseradish peroxidase (HRP) as a model glycoprotein, the proposed approach exhibited a wide linear range from 1 pg/mL to 100 ng/mL, with a low detection limit of 0.57 pg/mL. To the best of our knowledge, this is first report of a multiple signal amplification approach based on boronate-affinity molecularly imprinted polymer and SiO@Au/FcHT/MPBA, exhibiting greatly enhanced sensitivity for glycoprotein detection. Furthermore, the newly constructed BASA based glycoprotein sensor demonstrated HRP detection in real sample, such as human serum, suggesting its promising prospects in clinical diagnostics.

Ultrasensitive Bioaffinity Electrochemical Sensors: Advances and New Perspectives

“…Boronate affinity-based molecular imprinting is a new, facile, and reliable approach for imprinting sugars, small biological molecules, ribonucleo- tides, and glycoproteins because of its unique reversible covalent binding of boronate with cis-diol compounds [124]. Ultrasensitive electrochemical strategy was proposed for detection of glycoprotein based on boronate affinity sandwich assay and signal amplification with functionalized SiO 2 @Au nanocomposites [125]. In this approach, the AuNPs-GO was immobilized on the glassy carbon electrode (GCE) surface using chitosan, conjugated with 4-mercaptophenylboronic acid (MPBA) to capture target glycoprotein, and then fabricate the MIP film.…”

A novel Wulff‐type boronate acid‐functionalized magnetic metal‐organic framework imprinted polymer for specific recognition of glycoproteins under physiological pH

“…Protein glycosylation, a dynamic and ubiquitous protein posttranslational modifications, plays anextremely predominant role in numerous biological processes, including inter‐ and intracellular signal transduction, tumor immunology, cells migration, and protein folding [1‐2]. Moreover, research have shown that aberrant glycosylation is often accompanied by the occurrence of various physiological diseases, such as cancer, Alzheimer's cardiovascular disease, and Rheumatoid Arthritis [3‐5]. Hence, the specific analysis of glycoproteins is of great significance for the early diagnosis and real‐time monitoring of some diseases.…”

Molecularly imprinted polymeric shell coated monodisperse‐porous silica microspheres as a stationary phase for microfluidic boronate affinity chromatography

“…A general and simple approach was proposed for the preparation of molecularly imprinted magnetic nanoparticles by using glycoproteins as the templates . Boronate affinity sandwich assay on functionalized SiO 2 @Au nanocomposites was used for ultrasensitive electrochemical detection of glycoproteins . A molecularly imprinted polymer was evaluated for pH and temperature controlled sensing of glycopeptide based antibiotics .…”

“…The synthesis of molecularly imprinted sorbents by using boronate affinity interaction with the cis‐diol functionalized molecules has been usually performed by selecting nanoparticles as the supports . However, the imprinted nanoparticles are mostly suitable for the chromatographic applications performed based on biomagnetic separation conducted in batch fashion .…”