A spiral-electrospinning was used to mass-produce gelatin nanofibers with a content of chitosan oligosaccharide (COS) and alginate sodium (AS). Multiple jets were observed to form on the edges of the helix slice-spinneret simultaneously. Important electrospinning parameters, such as concentration of COS/gelatin aqueous solution, rotational velocity of spinneret and spinning distance, were examined to investigate the electrospinnability of COS/gelatin solution and the morphology of COS/gelatin nanofiber membranes. Due to the poor miscibility between COS and AS, COS/AS/gelatin nanofiber membranes were obtained from COS/gelatin solution and AS/gelatin solution by mixing electrospinning with multi-spinnerets. The novel needleless electrospinning not only avoided the possibility of nozzle-clogging, but also prepared COS/AS/gelatin nanofibers on a large scale for a wide variety of applications.