Development, simulation, and validation of microfluidic devices for biological applications: cell line culture, monoclonal antibody production, and drug screening in parasitic protozoa

Abstract

Microfluidics and lab-on-a-chip (LOC) devices enable the miniaturization and automation of complex biological processes at micro- and nanoscale levels. The objective of this thesis work is to develop, simulate, and validate microfluidic devices applied to three key areas: mammalian and stem cell culture, intensified production of monoclonal antibodies (mAbs), and automated drug screening in parasitic protozoa. A large-scale microfluidic bioreactor (LM-bioreactor) made of PDMS/glass was designed and implemented for the continuous production of a recombinant anti-hIFN-α2b antibody in CHO cells, achieving double the affinity constant (Ka) compared to conventional T-flask cultures. Additionally, a milli-bioreactor with a micropillar architecture was developed for passive bubble control during the culture of human induced pluripotent stem cells (hiPSCs), maintaining high expression levels of pluripotency markers Oct4, Nanog, and Sox2 for 5 days. Finally, a microfluidic dilutor device was used to evaluate 18 trypanocidal compounds against Trypanosoma cruzi via automated video tracking of motility (TR-KF software), identifying candidate 2161-X with high potency (IC50 = 0.2 µM) and selectivity index (SI = 115.6). Overall, these developments confirm low-cost flexographic mold-based microfluidics combined with CFD modeling as an efficient, scalable, and accessible platform for biotechnology and drug discovery.