High-affinity and specific recognition of human thyroid stimulating hormone (hTSH) by in vitro-selected 2'-amino-modified RNA

Abstract

RNA sequences containing 2'-amino pyrimidines that bind with high-affinity to human thyroid stimulating hormone (hTSH) were isolated from a random sequence library by an in vitro selection-amplification procedure. A representative RNA ligand (T-15) has an equilibrium dissociation constant (K(d)) of 2.5 nM for its interaction with hTSH and can discriminate between other members of the glycohormone family; no detectable binding was observed at low micromolar concentrations of hCG (human chorionic gonadotropin), while measured K(d) values for the interactions with hLH (human leutinizing hormone) and hFSH (human follicle stimulating hormone) were > 1 μM and ~ 0.2 μM, respectively. The detection of hTSH in a dot blot assay with radiolabeled T-15 RNA was demonstrated.