Campylobacter jejuni is one of the most common causes of traveller's diarrhoea and food poisoning, therefore development of a vaccine is important. Using biochemical fractionation and mass spectrometry analysis, we identified more than 110 surface polypeptides. Eight C. jejuni identified surface proteins were expressed in Escherichia coli and purified. Mice were immunized with different doses of these purified proteins and challenged orally with C. jejuni strains ML1 and ML53. The degree of protection of mice was tested by intestinal colonization. At least two groups of mice vaccinated with purified proteins clear the infection faster than control mice. Here, we present the use of a proteomics based approach for the identification of novel protein based C. jejuni vaccines for the first time.