| DYNWEB, s.r.o. | Liberec |
| E Linkx, a.s. | Prague |
| E+P Studio | Liberec |
| e-BAAN Net, s.r.o. | Prague |
| E-SOFT, spol. s r.o. | |
| E.C.Sys, s.r.o. | |
| e4t electronics for transportation, s.r.o. | Prague |
| E4YOU, spol. s r.o. | Prague |
| Eastburger.cz | Pardubice |
| eBRANA | Pardubice |
| Eccam, s.r.o. | Prague |
| ECOM, s.r.o. | Hradec Králové |
| Economia, a.s. | Prague |
| ECTACO Česká, s.r.o. | Prague |
| eD system, a.s. | |
| edgy.digital, s.r.o. | |
| eDisk, s.r.o. | South Moravian |
| Educasoft Solutions, s.r.o. | |
| EffiChem, s.r.o. | South Moravian |
| Eficia Praha, spol. s r.o. | Prague |
| eIdentity, a.s. | Prague |
| EKOL, spol. s r.o. | |
| EKOTRONIC, s.r.o. | Prague |
| Elanor | Prague |
| ELDATA | |
| ELDIS Pardubice, s.r.o. | |
| Electron Optical Design | South Moravian |
| ELEKTROSYSTEM, a.s. | |
| Eleveo | |
| Eline web, s.r.o. | Moravian-Silesian |
| Eline.cz | Moravian-Silesian |
| Elisoft | |
| Elix, spol. s.r.o. | Prague |
| ELMER software, s.r.o. | Prague |
| ELNET system, s.r.o. | Central Bohemian |
| Elsyst Engineering | South Moravian |
| Eltronic, s.r.o. | Liberec |
| ELVAC, a.s. | Moravian-Silesian |
| eM Client, s.r.o. | Prague |
| Emak.cz | Zlín |
| EmbedIT | Prague |
| EMEA, s.r.o. | Zlín |
| eMerite, s.r.o. | Prague |
| emocio, s.r.o. | Prague |
| EMP-Centauri, s.r.o. | Plzeň |
| enDetail, s.r.o. | South Moravian |
| Endora.cz | Liberec |
| Engeto, s.r.o. | South Moravian |
| Enter-Prise.net, s.r.o. | Prague |
| Envelon, s.r.o. | Prague |