Oriented surface imprinted 96-well microplate-based fluorescent biosensor for glycoprotein detection by boronate...

Area of Science:

• Biochemistry
• Analytical Chemistry
• Biomedical Engineering

Background:

• Glycoproteins are vital in biological processes and serve as key biomarkers and therapeutic targets in disease diagnosis.
• Detecting low-concentration glycoprotein biomarkers in complex biological samples is challenging due to interfering substances.

Purpose of the Study:

• To develop a specific, sensitive, and high-throughput fluorescent biosensor for glycoprotein determination in complex samples.
• To create an oriented surface-imprinted microplate-based fluorescent biosensor utilizing a boronate-affinity sandwich assay (BASA).

Main Methods:

• Development of a BASA employing boronate affinity-oriented surface-imprinted microplates and boronic acid-modified CdTe quantum dot (QD) fluorescence probes.
• Utilizing a sandwich assay format for specific binding of target glycoproteins.
• Implementing a "turn on" fluorescent detection mechanism.

Main Results:

• The biosensor demonstrated a wide linear range (1 ng/mL–10^5 ng/mL) and a low limit of detection (LOD) of 0.528 ng/mL for horseradish peroxidase (HRP).
• The "turn on" fluorescent sensor achieved at least three orders of magnitude higher sensitivity compared to traditional "turn off" sensors.
• Average recoveries ranged from 96.8% to 106.0% in urine samples, indicating high accuracy and reliability.

Conclusions:

• The developed BASA-based fluorescent biosensor offers a highly specific, sensitive, and high-throughput method for glycoprotein detection.
• This approach is suitable for analyzing glycoproteins in complex biological matrices, with potential applications in disease diagnosis and monitoring.