Earthworms possess antioxidant, antibacterial, antitumor, and hemolytic properties. To recognize the molecules responsible for various biological activities of earthworm’s coelomic fluid, a detailed knowledge about its protein contents is required. The aim of this study was to characterize the proteins present within the coelomic fluid of Eisenia foetida earthworm. Polyacrylamide-gel-electrophoresis (SDS-PAGE), and two-dimensional gel electrophoresis (2-DE) were carried out. Several proteins with molecular masses varying from 10 to 150 kDa were further separated from the coelomic fluid of Eisenia Foetida. The biological activities of the coelomic fluid could be mediated by these proteins.