AbstractThe production of polymer fibers from the combination of zein and PEO might have great potential in the field of biomaterial. Zein/PEO fibers were obtained in this work through solution electrospinning. An experimental design, 24‐1, was used for evaluating the influences of PEO content in the blend, distance from the needle tip to the collector, applied electric voltage and solution flow for average fiber diameter and relative‐yield process. Beyond this, the relationship between PEO content in the blend and the fiber properties were evaluated through FTIR, DSC, TG, tensile tests, and cytotoxic tests. The factor that exerts the greatest effect on the average fiber diameter response was the electrical voltage. The increase in PEO content in the blend decreased the thermal stability and increased the degree of the fibers' crystallinity. The mechanical tests showed that fibers with higher elongation were obtained at richer PEO blends. The fibers presented cytocompatible characteristics.