In this study, blend fibers composed of poly(vinyl alcohol) and poly(3,4-ethylenedioxythiophene):poly(styrene sulfonate) (PEDOT:PSS) were prepared via wet-spinning technology. Ethylene glycol (EG) vapor annealing was employed to improve the electrical conductivity and tensile properties of blend fibers. The effects of EG vapor annealing on structures and properties of blend fibers were investigated in detail by analyzing the changes in chemical constituent and structure, molecular structure, surface morphology, surface chemical composition, electrical conductivity, and tensile properties. FTIR spectroscopy indicates that EG vapor annealing does not change the chemical constituent and structure of blend fibers. Raman spectroscopy shows that vapor annealing leads to conformational changes of PEDOT chains from benzoid structure to quinoid structure. AFM and SEM images show that surface morphology of blend fibers become smoother after vapor annealing. XPS measurement shows that EG vapor annealing induces significant phase separation between PEDOT and PSS, forming an enriched PSS layer on the surface of blend fibers, thus leading to a thinner insulating PSS layer between PEDOT grains. This conformational change is beneficial to improve the electrical conductivity of blend fibers. The resultant blend fiber reached conductivity up to 20.4 S cm⁻¹. The mechanical properties of blend fibers were also improved by EG vapor annealing, with the Young’s modulus and tensile strength increasing from 3.6 GPa and 112 MPa to 4.4 GPa and 132.7 MPa, respectively.