High affinity ligands from in vitro selection: Complex targets

Abstract

Human red blood cell membranes were used as a model system to determine if the systematic evolution of ligands by exponential enrichment (SELEX) methodology, an in vitro protocol for isolating high-affinity oligonucleotides that bind specifically to virtually any single protein, could be used with a complex mixture of potential targets. Ligands to multiple targets were generated simultaneously during the selection process, and the binding affinities of these ligands for their targets are comparable to those found in similar experiments against pure targets. A secondary selection scheme, deconvolution-SELEX, facilitates rapid isolation of the ligands to targets of special interest within the mixture. SELEX provides high-affinity compounds for multiple targets in a mixture and might allow a means for dissecting complex biological systems.

A DNA Aptamer Prevents Influenza Infection by Blocking the Receptor Binding Region of the Viral Hemagglutinin

“…Such a library will generate more than 10 18 types of different ssDNAs, but only those that possess a fitting structure will hybridize with the peptide. Iteration of the selection procedure eventually leads to a small number of fitting aptamers (15). In the present study, it yielded just two novel oligonucleotides, A21 and A22, binding equally to the FIG.…”

A DNA aptamer population specifically detects Leishmania infantum H2A antigen

“…In Vitro Selection Procedure Iterative rounds of selection and amplification of ssDNA aptamers were performed as described previously by Morris et al 21 In brief, ssDNA oligonucleotides, designated as RND40, contained a central randomized region of 40 nucleotides flanked by two conserved 18-nucleotides regions in each end (5 0 -GCGGATGAAGACTGGTCT-40N-GTTGCTCGTATTTAGGGC-3 0 ). The ssDNA library was denatured at 901C for 10 min and then cooled on ice for 10 min.…”

Synthesis, Functionalization, and Design of Magnetic Nanoparticles for Theranostic Applications

“…[462,477] In recent years, the utility of aptamers has been enhanced through many techniques, in particular the one in which SELEX is performed with complex mixtures of biological materials and cell-SELEX. [498,533] By this technique aptamers which recognize specific cell lines are generated within an in vitro selection performed with whole live cells. [498,534] Finding more efficient aptamers for specific applications, they have also been selected in vivo by Mi et al in 2010.…”