Pre-absorbed immunoproteomics: a novel method for the detection of bacterial surface proteins

Bacterial surface proteins are often investigated as potential vaccine candidates and biomarkers of virulence. In this chapter, a novel method for identifying bacterial surface proteins is presented, which combines immunoproteomic with immunoserologic techniques. Immunoproteomics, involving the separation of proteins by two-dimensional electrophoresis (2-DE) and Western blotting, has become an increasingly popular method for identifying immunoreactive proteins. In conventional serological technique, cross-absorption is a powerful method used to minimize cross reaction during agglutination assays. The serum pre-absorption process in our method was developed from cross-absorption but modified to remove antibodies that recognized bacterial surface antigens, thereby generating pre-absorbed sera. These pre-absorbed sera were used in Western blotting after 2-DE to find bacterial surface protein antigens. This new method has been proven to be a useful tool for identifying surface proteins, and aid in the development of new vaccine subunits and disease diagnostics.