Development and analysis of glycoprotein based antiviral vaccines using Mass-Spectrometry

Human Cytomegalovirus (HCMV) is a b-herpesvirus widely spread over the whole globe, with a seropositivity varying from 50 to 100%. The infection by this virus can be very dangerous in immunocompromised patients (AIDS patients, transplant patients) and in infants, causing pneumonia, retinitis and life-long neurological damages. Currently no vaccines against HCMV are available; among promising candidate antigens for the development of a future vaccine against HCMV are two envelope glycoproteins: glycoprotein B (gB) and the pentameric complex gH/gL/UL128/UL130/UL131. These two antigens show several N- and O- glycosylation sites; a full study of glycans microheterogeneity and occupancy using LC-ESI-MS/MS is relevant for whole antigens characterization and immunological considerations. One of the aims of this project is to describe by combined enzymatic digestions the gB N-glycosylations sites using LC ESI-MS/MS. Furthermore, a released N-glycans analysis method will be developed using LC-FLR-ESI-MS 7 . Last but not least, another aim concerns the development of an O-glycosylation mapping method.