β-Conglutin dual aptamers binding distinct aptatopes.

Area of Science:

• Biochemistry
• Molecular Biology
• Immunotechnology

Background:

• A previous aptamer (β-CBA I) was selected against the anaphylactic allergen β-conglutin.
• The β-CBA I aptamer was truncated to an 11-mer, significantly improving its affinity.

Purpose of the Study:

• To select and characterize a second aptamer (β-CBA II) targeting a different site on β-conglutin.
• To develop and validate a dual-aptamer approach using both β-CBA I and β-CBA II for enhanced detection.

Main Methods:

• Aptamer selection and characterization against β-conglutin.
• Affinity validation using microscale thermophoresis and surface plasmon resonance.
• Dual-aptamer binding confirmation at different sites on the target allergen.

Main Results:

• A second aptamer, β-CBA II, was successfully selected with affinity comparable to the truncated β-CBA I.
• The dual-aptamer system demonstrated specific and simultaneous binding to distinct sites on β-conglutin.
• Independent laboratory validation confirmed the affinity and specificity of the aptamers.

Conclusions:

• The β-CBA II aptamer, in conjunction with β-CBA I, forms an effective dual-aptamer system.
• This dual-aptamer system can be utilized as a primary capturing and secondary signaling pair.
• The developed system holds potential for applications in enzyme-linked aptamer assays and aptasensors for allergen detection.