Multiplex Immunoassays Utilizing Differential Affinity Using Aptamers Generated by MARAS

Area of Science:

• Biotechnology
• Analytical Chemistry
• Molecular Diagnostics

Background:

• Accurate disease diagnosis relies on quantifying multiple biomarkers in patient serum samples.
• Existing multiplex immunoassay methods face challenges in sensitivity and specificity, necessitating novel approaches.

Purpose of the Study:

• To propose and demonstrate the feasibility of a novel method for multiplex immunoassays.
• To utilize the differential affinity of aptamers for simultaneous detection of multiple analytes.

Main Methods:

• Aptamers with varying affinities for multiple analytes were selected from a random oligonucleotide library using the Magnetic-field Assisted Aptamer Selection (MARAS) procedure under distinct magnetic field conditions.
• Multiplex detection was achieved by applying uniform magnetic field conditions to differentiate target analytes based on their aptamer affinities in patient serum samples.
• Assessing the recovery rates and non-specific binding in both buffer and serum matrices.

Main Results:

• The developed method demonstrated comparable recovery rates for spiked analytes in both buffer and complex biological matrices (serum).
• Minimal non-specific binding was observed, indicating high specificity for target molecules.
• The feasibility of using differential aptamer affinities for multiplex biomarker detection was successfully validated.

Conclusions:

• The novel aptamer-based multiplex immunoassay effectively utilizes differential binding affinities for accurate analyte differentiation.
• This method offers a promising platform for sensitive and specific disease diagnosis through multiplex biomarker quantification.
• The technique shows robustness in complex biological samples, paving the way for clinical applications.