There are several methods of preparing nanofiber yarns, but little work has been reported using a combination of electrospinning and solution blowing methods (i.e., electrostatic-solution blow spinning) to prepare nanofiber yarns. In this study, a response surface method (RSM) based on Box–Behnken design was used to investigate the relationship between the yarn parameters and yarn diameter, and the process was optimized. The yarn diameter was simulated by electrostatic voltage, air pressure, winding speed and funnel velocity. The correlation coefficients before and after adjustment were 0.97 and 0.95, respectively, through variance analysis, which verified the accuracy of the yarn diameter model. The results showed that the influence of each experimental factor on the yarn diameter was in descending order of winding speed, electrostatic voltage, airflow and funnel velocity, and there were interactions among the factors. The BDD response model was used to obtain the best experimental conditions, which provided a good theoretical reference for the preparation of polyacrylonitrile nanofibers. In addition, the effects of voltage and air pressure on the diameter of the nanofiber and yarn and the effects of the twist coefficient on the shape and mechanical properties of the yarn were studied. The results showed that with the increase of voltage and air pressure, the diameter of the nanofiber decreased, and the yarn diameter decreased first and then increased. When the twist factor was 100, the twist angle of the yarn was 49.91°, the breaking strength was 12.92 MPa and the elongation at break was 81.34%.