Morphological and cytoskeletal changes caused by non-membrane damaging cytotoxin of Vibrio cholerae on Int 407 and HeLa cells

End-group mediated conjugation of bacterial polysaccharides (PSs) to carrier proteins containing T-helper cell epitopes renders such polysaccharides immunogenic also in young infants. Optimal construction of such conjugate vaccines requires fragmentation of the PS prior to the coupling reaction. In the present study a general simple and inexpensive method for the fragmentation of PSs is presented. It is based on the irradiation of isolated PSs in an electron beam accelerator. Exposure of isolated pneumococcal capsular polysaccharides (PnPSs) to ionizing radiation resulted in their partial depolymerization in a radiation dose-dependent manner. Radiation, unlike sonication, generated PnPS fragments of molecular size lower than 50 kDa and as small as 1.5 kDa when high radiation doses were used. These PnPS fragments have terminal reducing groups that can be easily used for chemical activation and subsequent coupling to any chosen carrier protein. The radiation-produced PnPS fragments retained their antigenic epitopes, when compared to native, full-size PnPSs as determined by enzyme-linked immunoassay.