An Enzyme-Linked Aptamer Sorbent Assay to Evaluate Aptamer Binding.

Area of Science:

• Biochemistry
• Molecular Biology
• Analytical Chemistry

Background:

• Nucleic acid aptamers are emerging as powerful tools in molecular detection, offering advantages over traditional ligands like antibodies.
• Confirming specific target molecule binding is crucial after aptamer generation.
• The enzyme-linked aptamer sorbent assay (ELASA) is a rapid method for screening aptamer-protein interactions.

Purpose of the Study:

• To describe and validate an enzyme-linked aptamer sorbent assay (ELASA) for evaluating aptamer binding.
• To compare the binding capabilities of single-stranded DNA (ssDNA) aptamers against different strains of human norovirus capsid proteins.

Main Methods:

• Directly plating protein targets (human norovirus capsids) onto a polystyrene surface.
• Utilizing functionalized (biotinylated) ssDNA aptamers to assess binding to the immobilized protein targets.
• Employing an enzyme conjugate that recognizes the aptamers to detect binding events.

Main Results:

• The study successfully designed and implemented an ELASA protocol.
• The assay allowed for the evaluation and comparison of ssDNA aptamer binding affinities.
• Binding of aptamers to different human norovirus strains was characterized.

Conclusions:

• The described ELASA is an effective method for screening and characterizing aptamer-protein interactions.
• This assay facilitates the comparison of aptamer binding to various target strains.
• The findings support the utility of aptamers in detecting specific viral targets like human norovirus.