In addition to its natural ligand, the receptor for erythropoietin can be activated by small peptides known as erythropoietin mimetic peptides (EMPs). Although EMPs are less potent than the natural ligand, EMP dimers, consisting of two EMPs joined via a linker, have been shown to exhibit significantly improved activity compared to the corresponding monomers, with potency approaching that of the native hormone. In this study, we used a panel of novel EMP dimers to explore the effects of linker length and EMP attachment site on potency. The EC50 values obtained in an EPO-dependent proliferation assay indicated that, as has been shown with similar molecules, EMP dimerization can lead to increases in potency of more than 2 orders of magnitude. We found that both C-terminal and N-terminal attachment of the linker to EMP was tolerated, and that, with the exception of the shortest linker, all of the linker lengths tested provided a similar increase in potency. In follow-up work devised to explore the potential benefit of contacting additional cell surface EPO receptors, we designed a tetrameric template consisting of lysine-based dimers joined via commercial PEG linkers of various molecular weights. Evaluation of the resulting molecules indicated a clear effect of PEG linker size on activity, while the "dimer of dimer" with the shortest linker exhibited 10-fold lower potency than the corresponding dimer, the longest tetramer increased potency by fivefold. We discuss the implications of these results for the further development of EMP multimers.