Botulism is a neuroparalytic disease caused by botulinum neurotoxins (BoNTs) and has been reported as a potential bio warfare agent due to its extreme toxicity (~100 billion times more toxic than cyanide). BoNTs is produced by Gram positive; an obligate anaerobic and endospore forming bacteria called Clostridium botulinum. There is no licensed vaccine available commercially for the treatment of botulism. An immunoproteomic-based approach was used to screen the immunogenic proteins of C. botulinum type B. The identity of immunogenic proteins is important to develop the vaccine candidates and diagnostic markers against the botulism. Using a proteomics-based approach, whole cell proteome were separated by two-dimensional gel electrophoresis and immunoreactive proteins were revealed by reaction with antisera against whole cell proteins of C. botulinum type B. The identity of immunoreactive proteins were recognized by mass spectrometry. Fifty six pre dominant proteins were identified in total. Out of total proteins, 13- proteins were identified as immunogenic. These proteins may be potential vaccine / diagnostic candidate molecule against botulism.