Descripción del proyecto
THE GOAL OF THE PROJECT IS TO DEVELOP A VERSATILE, OPTIMIZED AND REPRODUCIBLE BIOPROCESS PLATFORM TO OBTAIN NEW VACCINE CANDIDATES, THAT CAN BE FURTHER ADAPTED FOR THE PRODUCTION OF A MYRIAD OF VACCINE PRODUCTS, AND SERVE TO EVALUATE THEIR EFFICACY, INDUSTRIAL POTENTIAL AND TECHNICAL AND ECONOMICAL FEASIBILITY. THE MOST IMPORTANT ASPECT OF THESE NEW TYPES OF VACCINES, IN CONTRAST TO THE CLASSICAL ONES, IS THEIR BIOSAFETY, WHILE MAINTAINING THE EFFICIENCY FOUND IN THE MOST CONVENTIONAL LIFE-ATTENUATED VACCINES. THE PROJECT FOCUSES ON THE DEVELOPMENT OF A BIOPROCESS BASED ON ANIMAL CELL CULTURE. PROPER BIOPROCESSING STRATEGIES NEED TO BE ESTABLISHED AND OPTIMIZED FOR THE PRODUCTION OF THESE COMPOUNDS. THE DEVELOPED SYSTEM SHOULD BE COMPLIANT WITH GMP STANDARDS, AND FOR THIS OBJECTIVE THE USE OF SINGLE USE DISPOSABLE ELEMENTS IN THE COMPLETE PROCESS (UP-STREAM, BIOREACTOR AND DOWN-STREAM) IS PROPOSED AS A NEW ENABLING TECHNOLOGY (ALTHOUGH STILL LACKING OF ENGINEERING DEVELOPMENT) AND WILL BE IMPLEMENTED WITH THE CORRESPONDING MONITORING AND CONTROL ELEMENTS. TO BRIDGE THE UNDERSTANDING AND CHARACTERIZATION OF THE BIOLOGICAL ELEMENTS WITH THE BIOPROCESS DEFINITION, OPERATION, CONTROL AND OPTIMIZE PRODUCTIVITY, MATHEMATICAL MODELLING PLAYS A CENTRAL ROLE, INCLUDING METABOLIC FLUX ANALYSIS. THE COMPLETE CHARACTERIZATION OF THE COMPOUNDS PRODUCED IN THIS PLATFORM IS ALSO VERY IMPORTANT, FOR INSTANCE IN TERMS OF PRODUCT CONSISTENCY AND IMMUNOGENIC RESPONSE GENERATED IN ANIMAL MODELS, AS A PROOF OF CONCEPT OF THEIR EFFICIENCY. THE TECHNICAL AND ECONOMICAL VIABILITY OF THE PROCESSES DEVELOPED WILL ALSO BE STUDIED, CONSIDERING THE ASPECTS ASSOCIATED TO THE SCALE-UP OF THE PROCESS TO THE REQUIRED INDUSTRIAL SCALE ACCORDING TO THE PRODUCTION NEEDS. TWO DIFFERENT TYPES OF NEW GENERATION VACCINES WILL BE STUDIED, WITH APPLICATIONS IN ANIMAL AND HUMAN HEALTH. IN BOTH CASES, THE PRODUCTION WILL BE BASED ON ANIMAL CELL CULTURE DUE TO THEIR SUPERIOR CAPACITY TO OBTAIN FINAL PRODUCTS WITH CORRECT GLYCOSYLATION PATTERNS AND THEREFORE BIOLOGICAL ACTIVITY. THE MAMMALIAN CELL LINE PROPOSED IS THE HEK 293, WIDELY USED IN THE FIELD OF VACCINE PRODUCTION, WORKING IN SUSPENSION CULTURE. THE HUMAN HEALTH APPLICATION FOCUSES ON THE PRODUCTION OF VIRUS-LIKE PARTICLES (VLP)-BASED VACCINES. EXPRESSION OF GENES FROM HIV VIRUS, PARTICULARLY GAG, ENABLE THE FORMATION OF A VLP STRUCTURE THAT CAN BE FURTHER MODIFIED TO EXPOSE A SPECIFIC ANTIGEN ON ITS SURFACE REQUIRED TO BUILD THE VACCINE PARTICLE. ONE OF THE MAIN ADVANTAGES OF SUCH TYPE OF PARTICLES IS THE ABSENCE OF VIRAL GENOME AND THEREFORE VLP-BASED VACCINES ARE NON-INFECTIVE BY NATURE. THE ANIMAL HEALTH APPLICATION WILL MAKE USE OF ADENOVIRUS VECTORS (ADV) CODING FOR SPECIFIC ANTIGENS. THIS STRATEGY RELIES ON THE CELL MACHINERY OF THE HOST TO PRODUCE IN VIVO THE REQUIRED ANTIGENS, FOLLOWING THE INTRODUCTION OF THE FOREIGN GENES BY THE VIRAL VECTOR. THE HIGHLY SOPHISTICATED MACHINERY OF THE ADV FACILITATES AN EFFICIENT INTRODUCTION OF THE GENE OF INTEREST INTO THE HOST CELLS. A MORE ADVANCED POSSIBILITY WILL BE THE MODIFICATION OF THE ADV CAPSID BY THE INSERTION OF THE IMMUNOGENIC ANTIGEN. IN THIS CASE THE VACCINE WILL CONSIST ON THE RECOMBINANT ADV DIRECTLY ADMINISTRATED TO THE ANIMALS. THIS PART OF THE WORK WILL FOCUS ON THE DEVELOPMENT OF A CANDIDATE VACCINE BASED ON ADV TECHNOLOGY FOR THE PROPHYLAXIS OF PORCINE CIRCOVIRUS INFECTION.