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A Universal Boronate-Affinity Crosslinking-Amplified Dynamic Light Scattering Immunoassay for Point-of-Care
Jing Chen1, Liangwen Hao1, Jiaqi Hu1
1State Key Laboratory of Food Science and Technology, School of Food Science and Technology, Nanchang University, Nanchang, 330047, China.
Insights
This study introduces a new assay for detecting glycoproteins using dynamic light scattering (DLS) and boronate-affinity crosslinking. The method enables rapid, ultrasensitive glycoprotein detection at the point-of-care.
Area of Science:
- Biomedical Engineering
- Analytical Chemistry
- Nanotechnology
Background:
- Glycoprotein detection is crucial for diagnosing various diseases.
- Existing methods for glycoprotein analysis can be complex and time-consuming.
- Point-of-care diagnostics require sensitive, rapid, and user-friendly assays.
Purpose of the Study:
- To develop a universal boronate-affinity crosslinking-amplified dynamic light scattering (DLS) immunoassay.
- To enable sensitive and rapid glycoprotein detection in complex samples at the point-of-care (POC).
Main Methods:
- Utilized antibody-coated magnetic nanoparticles (MNP@mAb) for glycoprotein capture and DLS signal transduction.
- Employed phenylboronic acid-based materials as crosslinking amplifiers, leveraging selective binding to cis-diol-containing glycoproteins.
- Implemented target-induced crosslinking to amplify the DLS signal based on changes in average nanoparticle size.
Main Results:
- Achieved ultrasensitive quantitative assay of glycoproteins at the femtomolar (fM) level.
- Demonstrated rapid detection within 15 minutes.
- The assay showed high sensitivity and speed due to multivalent binding and fast boronate-affinity reactions.
Conclusions:
- The developed DLS immunoassay offers a versatile and promising strategy for glycoprotein detection.
- This approach extends the utility of DLS for field or point-of-care applications.
- The boronate-affinity crosslinking amplification significantly enhances sensitivity and speed for glycoprotein analysis.
Abstract:
Herein, we report a universal boronate-affinity crosslinking-amplified dynamic light scattering (DLS) immunoassay for point-of-care (POC) glycoprotein detection in complex samples. This enhanced DLS immunoassay consists of two elements, i.e., antibody-coated magnetic nanoparticles (MNP@mAb) for target capture and DLS signal transduction, and phenylboronic acid-based boronate-affinity materials as crosslinking amplifiers. Upon the addition of targets, glycoproteins are first captured by MNP@mAb and amplified by target-induced crosslinking stemming from the selective binding between the boronic acid ligand and cis-diol-containing glycoprotein, thereby resulting in a remarkably increased DLS signal in the average nanoparticle size. Benefiting from the multivalent binding and fast boronate-affinity reaction between glycoproteins and crosslinkers, the proposed immunosensing strategy has achieved the ultrasensitive and rapid quantitative assay of glycoproteins at the fM level within 15 min. Overall, this work provides a promising and versatile design strategy for extending the DLS technique to detect glycoproteins even in the field or at POC.
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