Particular thanks go to the child group management and staff and the parents who participated. “
“Foot-and-mouth disease (FMD) is an economically important viral disease that affects animals such as cattle, swine and sheep with a potential for rapid spread. The causative agent, FMD virus (FMDV), is a positive-stranded RNA virus enclosed by an icosahedral capsid. Intact (infectious) FMDV particles sediment at 146S in sucrose gradients. They are composed of 60 copies of VP1, VP2, VP3 and VP4 each and the RNA molecule [1]. Specific degradation products of such virions can be generated by mild acid treatment or heating to 56 °C. These 12S

particles consist of 5 copies of VP1, VP2 and VP3 each and lack VP4 [2]. Seven antigenically distinct serotypes of FMDV have been identified: O, A, C, Asia 1, SAT1, SAT2 and SAT3 [3]. Conventional FMD

AZD6244 vaccines are based on virus that is cultured using baby hamster kidney (BHK)-21 cells, inactivated by binary ethyleneimine (BEI) treatment, concentrated and formulated with a suitable adjuvant. Such FMD vaccines are unstable as measured by potency tests or serology [4] and [5]. The molecular basis for this decrease in FMDV immunogenicity is unclear. Proteolysis of FMDV antigens has been detected during prolonged storage at 4 °C [6] and [7]. Dissociation of 146S particles into 12S particles could also be involved [8]. Finally, specific chemical modifications such as deamidation or oxidation of specific amino acids

could also negatively affect vaccine efficacy, as was TAM Receptor inhibitor demonstrated for several other vaccine antigens [9] and [10]. However, chemical modification of FMDV antigen has never been analysed. In this study we used surface-enhanced laser desorption ionization-time of flight-mass spectrometry (SELDI-TOF-MS) for profiling of FMDV antigen. This method uses several ProteinChip arrays for immobilization of proteins on various chromatographic surfaces dependent on their physicochemical characteristics. Antibodies can also be covalently coupled to activated surfaces of particular ProteinChip arrays for specific immunocapture of the target antigen ADP ribosylation factor from complex samples. The noncovalently bound antigens are then analysed by TOF-MS. Advantages of SELDI-TOF-MS as compared to Western blotting or 2D SDS-PAGE are higher sensitivity (at least at lower molecular mass), low sample volume, ease-of-use, speed and high reproducibility [11]. SELDI-TOF-MS is often used for proteomic analysis of complex samples such as blood or urine. However, it is also suitable for other applications such as expression optimization and purification process development [12]. Here we have used SELDI-TOF-MS for characterization of FMDV antigen during various stages of vaccine production.