Metal nanoparticles exhibit unique size- and spatial organization-dependent physical and chemical properties, and have a wide range of applications in various areas including single electron devices, chemical catalysts and biomedicines. In this paper, chains of palladium nanoparticles were obtained by incubating aged sodium tetrachloropalladate(II) with glucagon fibrils pre-deposited on a solid surface. AFM height profiles showed that the size of the palladium nanoparticles within the chains could be fine tuned in the range of ∼2 to 16 nm as a function of the concentration of the sodium tetrachloropalladate(II). Moreover, the coverage of the palladium nanoparticles along the fibrils was controlled simply by varying the incubation time. This method provides a facile approach for the construction of a palladium nanoparticle ensemble on biotemplates.