Solid phase peptide synthesis is a method used to generate peptides through the sequential addition of amino acids attached to an immobilized resin. Due to the procedure followed, excess reagents and byproducts are removed while synthesis is ongoing, resulting in a final product that is reproducibly obtained at high purity. Herein, we describe the synthetic procedure of peptides derived from the pleotropic protein macrophage migration inhibitory factor (MIF). Notably, this protocol is valid for the generation of peptides regardless of their sequence and length. Whereas MIF has diverse activities and protein partners, the generation of thoughtfully designed MIF peptides may have applications in drug discovery and structural biology.