Tyramide signal amplification for a highly sensitive multiplex immunoassay based on encoded hydrogel microparticles

Area of Science:

• Biomedical research
• Immunology
• Assay development

Background:

• Protein quantification is vital for understanding diseases.
• Conventional immunoassays lack sensitivity, dynamic range, and multiplexing capability for clinical samples.

Purpose of the Study:

• Develop a highly sensitive multiplex immunoassay.
• Improve detection limits for proteins in biomedical research and diagnostics.

Main Methods:

• Utilized encoded hydrogel microparticles (MPs) with tyramide signal amplification (TSA).
• Optimized detection antibody concentration, biotinyl tyramide concentration, and TSA reaction time.
• Blocked unreacted acrylate double bonds to reduce non-specific binding and increase signal-to-noise ratio.

Main Results:

• Achieved a limit of detection as low as 58 fg mL⁻¹.
• Demonstrated a 10-fold enhancement in assay sensitivity compared to previous methods.
• Successfully multiplexed five cytokines (IL-4, IL-5, IL-6, IL-9, IL-17) in human serum without cross-reactivity.

Conclusions:

• The developed immunoassay offers significantly enhanced sensitivity and multiplexing capacity.
• This platform enables detection of low-abundance proteins in clinical samples, advancing diagnostics.
• The approach holds promise for biomedical research and clinical applications requiring precise protein quantification.