Evaluation of sensing ability of aptamers towards β-casomorphin-7 (BCM-7) peptide in different assay formats

Abstract

Aptamers are useful ligand molecules for the recognition of target molecules. Aptamers, as well as target molecules, are modified for their use in developing assay protocols for targets. Assay format uses gold nanoparticles or magnetic beads or 96-well plate or membrane surface for interaction between aptamer and target molecules. The present investigation is aimed to evaluate the recognition potential of modified aptamers towards β-casomorphin-7 (BCM-7) peptide. Aptamers (Seq.7 and Seq.U5) selected in previous work for BCM-7 as well as their variable regions and selected truncated segments of variable regions were evaluated for recognition of BCM-7. Aptamers or biotinylated aptamers or amino group attached to aptamer and BCM-7 or biotinylated BCM-7 or BCM-7-peroxidase were used in different assay geometry using streptavidin-coated gold nanoparticle or streptavidin-coated magnetic beads or streptavidin-coated 96-well plate or nitrocellulose & nylon membranes. In different assay formats, biotinylated full-length aptamers (Seq.7 and Seq.U5) as well as biotinylated variable region of aptamer Seq.U5 (Seq.U5v) recognized BCM-7. However, truncated aptamers from Seq.U5v failed to recognize BCM-7. Assay formats using aptamer-BCM-7 interaction can be adapted for studying aptamer-target recognition. Seq.7 and Seq.U5 are promising aptamers for developing protocols in different assay geometry for measuring BCM-7 in the urine sample.

URN:NBN:sciencein.nanomed.2023.v3.554