Cross-Reactive Arrays Based on Three-Way Junctions

• Published in: Journal of the American Chemical Society Vol. 125; no. 20; pp. 6085 - 6089
• Main Authors: Stojanović, Milan N, Green, Eric G, Semova, Stanka, Nikić, Dragan B, Landry, Donald W
• Format: Journal Article
• Language: English
• Published: Washington, DC American Chemical Society 21-05-2003
• Subjects: Analytical chemistry; Biological and medical sciences; Biosensing Techniques - methods; Biosensors; Biotechnology; Chemistry; Cocaine - analysis; Cross Reactions; Exact sciences and technology; Fluorescent Dyes - chemical synthesis; Fluorescent Dyes - chemistry; Fundamental and applied biological sciences. Psychology; General, instrumentation; Hydrophobic and Hydrophilic Interactions; Ligands; Methods. Procedures. Technologies; Nucleic Acid Conformation; Oligonucleotides - chemical synthesis; Oligonucleotides - chemistry; Phosphorous Acids - chemical synthesis; Phosphorous Acids - chemistry; Structure-Activity Relationship; Various methods and equipments; Performance evaluation; Fluorescence detector; Binding capacity; Double helical structure; Biosensor; Molecular recognition; Hydrophobic compound; Oligonucleotide; Chemical sensor
• ISSN: 0002-7863; 1520-5126
• DOI: 10.1021/ja0289550

We report herein a novel system for the parallel processing of molecular recognition events utilizing arrays of oligonucleotide-based fluorescent sensors to characterize hydrophobic molecules in solution. The binding domains of the sensors were based on three-way junctions that utilize double helical stems as framework regions to reliably fold regardless of variations in or around the binding domain. A reporting domain was introduced by the specific substitution of a single phosphodiester group with a phosphorothioate, followed by selective functionalization with a fluorophore. The sensors were organized into cross-reactive arrays to yield characteristic fingerprints for samples containing hydrophobic molecules. The fingerprints can be used to characterize steroids in solution, including complex biologically important fluids. Arrays have the potential for clinical applications such as the detection of gross errors in steroidogenesis.