We successfully developed DNA aptamers which can specifically recognize single-chain variable fragments (scFvs). These aptamers can contribute to specific and universal detection of scFvs, which will be utilized for the future therapeutic scFv continuous sensing.<br/>ScFvs are one of the most widely used engineered fragment antibodies, fabricated by joining together the variable heavy and light chain of an antibody via a peptide linker. Because they can be produced by cost-effective bacterial expression systems and engineered easily, scFvs are now utilized for therapeutic reagents. However, it is generally difficult to detect scFv specifically suitable for the pharmacokinetic analyses.<br/>Here, we aimed to obtain scFv-specific DNA aptamers, as the future biosensing molecule to detect therapeutic scFv. Aptamers were screened by Systematic Evolution of Ligands by Exponential enrichment (SELEX) with combining competitive selection using scFv, and concentrated sequences and sequences with common motifs were selected as candidates. For further selection, evaluation of binding to scFv was performed by Enzyme-linked oligonucleotide assay (ELONA). Thus, obtained aptamers can specifically recognize and bind to scFv, which were confirmed by dot blotting and Biolayer Interferometry (BLI). Furthermore, these aptamers revealed that they did not bind to Fv harboring the same CDR region of scFv used for SELEX, and also showed the binding ability to other scFvs with different CDR region. We will present the construction of aptamer sensor for scFv monitoring.

Development of anti-single-chain variable fragments (scFv) DNA aptamers for the therapeutic scFv sensing

Fall 2022 abstracts, posters, and presentations will launch in mid-September.

ACS Fall 2022

The official archive and repository for all ACS Meetings & Expositions abstracts and select posters or presentations.