Acetone-butanol-ethanol (ABE) fermentation using Clostridium acetobutylicum is a process that can be used to produce butanol, which can be utilized as an alternative to petroleum-based fuels. Immobilization of the bacteria using three different fibrous materials was studied in order to see how to improve the ABE fermentation process. The results were compared to those of non-immobilized bacteria. Modal and charcoal fibers had OD levels below one at 72 h with the butanol concentration reaching 11.0 ± 0.5 and 10.7 ± 0.6 g/L, respectively, each of which were close to the free cell concentration at 11.1 ± 0.4 g/L. This suggests that bacteria can be efficiently immobilized in these fibrous materials. Although an extended lag phase was found in the fermentation time course, this can be easily solved by pre-treating fibrous materials with 3.5% HCl for 12 h. From comparisons with previous studies, data in this study suggests that a hydrophilic surface facilitates the adsorption of C. acetobutylicum.