An end-to-end assembly of spherical Ag nanoparticles takes place in the presence of biotin to form long fiberlike microstructures. These microstructures are about 4 μm long with a thickness of 1 μm, obtained from SEM studies. TEM studies showed the presence of spherical silver nanoparticles having an average size of 20 nm. ATR–FTIR studies revealed that silver ions interact with biotin involving the carboxylate group. A weak binding of the silver particles with the thioether and ureido groups helps in connecting the Ag nanoparticles to form long fiberlike structures. Elucidation of the mechanism of formation of the spherical Ag clusters was done by pulse radiolysis.