Combining the Benefits of Biotin–Streptavidin Aptamer Immobilization with the Versatility of Ni-NTA Regeneration Strategies for SPR

• DOI: 10.3390/s24092805
• Publication Date: 2024-04-29T12:49:24Z
• Authors: Eliza K. Hanson; Rebecca J. Whelan
• Keywords: Streptavidin; Aptamer; Nitrilotriacetic acid

The high affinity of the biotin–streptavidin interaction has made this non-covalent coupling an indispensable strategy for immobilization and enrichment biomolecular reagents. However, irreversible nature bond renders surfaces functionalized using permanently modified not amenable to regeneration strategies that could increase assay reusability throughput. To utility biotinylated targets, we here introduce a method reversibly immobilizing thrombin-binding aptamers onto Ni-nitrilotriacetic acid (Ni-NTA) sensor chip 6xHis-tagged streptavidin as regenerable capture ligand. This approach enabled reproducible measurements aptamer–protein in surface plasmon resonance assay. immobilized aptamer was stable during five experiments over two days, despite reversible attachment 6xHis-streptavidin Ni-NTA surface. In addition, demonstrate reproducibility assays performed it. Finally, verify specificity biotin tag–streptavidin assess efficiency straightforward regenerate reuse described will allow researchers leverage versatility stability while increasing throughput improving efficiency.