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Technological platform for production of monoclonal antibodies fragments using recombinant Escherichia coli


Monoclonal antibodies (mAbs) are a class of biotherapeutic against a range of major diseases, making a current market of $ 125 billion by 2020. The industrial production system of choice is the submerged cultivation of animal cells in large scale bioreactor. The structural complexity of mAbs, low cell growth velocity, use of complex culture media, sensitivity to temperature variations and concentration gradients at scaling up, has attracted interest in the production and use of Fab (antigen) antibody fragments. antibody Fragment) and scFv (single-chain variable Fragment) in recombinant E. coli systems due to their lower production complexity and potential lower production cost. Stx1 and Stx2 toxins are responsible for causing serious pathologies such as hemorrhagic colitis, uremic hemolytic and renal failure. These conditions are caused by infection of enteropathogenic bacteria with consequent production of Stx1 and Stx2, and there is currently no therapy for their effective combat. Recently, recombinant E coli expressing scFv and Fab fragments against Stx1 and Stx2 toxins were constructed and demonstrating immunogenic pattern in vivo, showing potential therapeutic and diagnostic effect against Stx1 and Stx2 (Luz et al. 2015,2018). The aim of this project is to develop a high cell density production process of recombinant Escherichia coli for the production of Fab and scFv antibody fragments against Stx1 and Stx2 toxins, using these recombinant strains, outlining the main stages of fragment separation and purification, and performing preliminary economic analysis of the production process to identify their bottlenecks. (AU)

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