【Abstract】 Objective To investigate the components of 68kD inner ear antigens inducing autoimmune Meniere’s disease (AIMD) in Guinea pigs. Methods Inner ear antigens of Guinea pigs were separated by two-dimensional polyacrylamide gel electrophoresis (2-DE). The 68kD protein spots were chosen by ImageMaster software and mass spectrometry was used to test these protein spots. The Swiss-prot database was searched with Mascot software and then the PathwayStudio software was used to carry out the bioinformatics analysis for proteins searched. Western blotting was performed to prove selective proteins. Results 2-DE patterns with higher resolution and reproducibility was obtained. It was found that a total of 22 kinds of protein spots contained in 68kD antigens were acquired by ImageMaster software analysis, and 13 kinds of protein were identified with mass spectrometry and Swiss-prot database searching. These proteins were related to immunity, apoptosis, differentiation and other functions respectively, in which vimentin may be the main proteins in 68kD inner ear antigens for inducing AIMD in Guinea pigs according to assessment from relative molecular mass, Mascot score and protein functions. Conclusions 2-DE pattern of 68kD proteins of guinea pig inner ear antigens that were successfully was obtained, and the total of protein spots were identified with mass-spectrometric technique. Vimentin may be the main proteins in 68kD antigens for inducing AIMD in guinea pigs.