Desenvolvimento de processos em perfusão a baixas taxas de alimentação para a produção de anticorpos monoclonais recombinantes com células CHO

Abstract

Monoclonal antibodies are the most approved biopharmaceuticals for human therapy. A higher demand for those molecules and the competition with biosimilars have driven biomanufacturing companies to establish more efficient production platforms. Continuous processes have been proposed due to smaller footprint, more compact systems, higher productivities and a more consistent product quality. However, a limited number of cell retention devices and the high quantities of medium and supernatant hamper a widespread adoption of perfusion over fed-batch processes. In this work, cellspecific perfusion rates (CSPR) lower than 20 pL/cell/day were applied in perfusion processes with two Chinese hamster ovary (CHO) cell lines. Cell retention was performed with three different devices: alternating tangential flow (ATF) and cell settler, which are widely used in industry; and a hydrocyclone, which had been tested with relatively lowdensity cell suspensions. For the first time, viable cell densities between 20x106 and 50x106 cell/mL were achieved in a perfusion with a hydrocyclone operated at pressure drops higher than 1 bar. Cultivations were maintained up to 50 days in lab-scale bioreactors and in a 50-L single-use bioreactor bag. A maximum cell density of 130x106 cell/mL was achieved at 19 pL/cell/day with ATF. As expected for membraneless equipment, product retention in the bioreactor was not observed using cell settler and hydrocyclone. The reduction in CSPR provided higher product titers, even though a decrease in growth and productivity rates was observed.