Tutorial: design and fabrication of nanoparticle-based lateral-flow immunoassays

Area of Science:

• Biosensors and Diagnostics
• Point-of-Care Testing
• Biotechnology

Background:

• Lateral-flow assays (LFAs) are widely used, rapid, and cost-effective biosensors.
• Their operation relies on capillary flow for detecting diverse targets in various sample types.
• Developing LFAs involves optimizing multiple interconnected parameters, which can be challenging for new researchers.

Purpose of the Study:

• To provide fundamental knowledge for understanding LFA principles and design.
• To offer a universal roadmap for LFA development, adaptable to specific applications.
• To present a practical, step-by-step protocol for assembling and operating an LFA for human IgG detection.

Main Methods:

• A tutorial-based approach explaining LFA principles and design considerations.
• A structured roadmap for LFA development applicable across different targets.
• A detailed example procedure for constructing and using a lateral flow strip.
• An extensive troubleshooting guide for common LFA development and usage issues.

Main Results:

• Readers gain foundational knowledge for informed LFA design and fabrication decisions.
• A clear development pathway is established, simplifying the process for new users.
• A functional LFA protocol for human IgG detection is demonstrated.
• Common challenges in LFA development and use are addressed.

Conclusions:

• This tutorial empowers researchers to independently develop and optimize LFAs.
• The adaptable protocol facilitates cost-efficient detection of a wide range of analytes.
• LFAs offer a versatile platform for point-of-care and field-based diagnostics.