Production of monoclonal antibodies in microfluidic devices

Abstract

Herein, a microfluidic device with cistern design for cultivation of adherent eukaryotic cells for the production of recombinant proteins is presented. The geometric configuration of the microchannels in the device provided laminar flow with reduced velocity profiles in the cisterns, resulting in an adequate microenvironment for long-term adherent cell growth with passive pumping flow cycles of 24 hours. CHO-ahIFNα2b and HEK-ahIFNα2b adherent cell lines expressing a novel anti-hIFN-α2b recombinant monoclonal antibody (MAb) for the treatment of systemic lupus erythematosus were cultured on the surface of PDMS/glass microchannels coated with poly-d-lysine. A 24 day culture of CHO-ahIFNα2b cells resulted in MAb concentrations up to 166.4 μg mL per day. The productivity of CHO-ahIFNα2b and HEK-ahIFNα2b cell lines was higher in the microdevice compared to that obtained using the adherent cell culture method (T-flask), with a 5.89- and 7.31-fold increase, respectively. Moreover, biological analysis of the MAbs produced in the microdevice showed no significant differences in the neutralizing antiproliferative activity of the hIFN-α2b or the cytokine cell signaling compared to the MAbs produced with cell adherent methods. These results suggest that this microfluidic device is suitable for long-term culture of mammalian cells and can improve the productivity of cells expressing recombinant MAbs with potential for therapeutic use without affecting the quality attributes of the product.

Large Area Microfluidic Bioreactor for Production of Recombinant Protein

“…In addition, >900,000 freedom elements (average element quality of >0.6) were included for the construction of the mesh density. The methodology was modified from [ 25 ].…”

“…The Chinese hamster ovary CHO-ahIFN-α2b adherent cell line was used [ 25 ] to evaluate the production of recombinant antibodies in the MM bioreactor. This cell line expresses the anti-hIFN-α2b recombinant scFv-Fc and was obtained from Attallah et al, at Centro Biotecnológico del Litoral (UNL/CONICET) as previously described [ 34 ].…”

“…There are few studies on the use of LOC-type devices to produce recombinant proteins [ 24 ]. In previous work, we have demonstrated the feasibility of the production of an anti-hIFN-α2b recombinant mAb with Chinese hamster ovary, CHO, and human embryonic kidney HEK-293 cells in standard-size microfluidic devices with a final volume of 32.22 µL [ 25 ]. It was possible to increase the concentration of the antibodies up to 7.3 times higher (1120 pg of mAb per cell) than that obtained by standard culture methodologies (using T-flasks).…”

Application of Microfluidic Systems for Breast Cancer Research

“…Biological therapeutics such as MAbs, CAR-T, vaccines, and virotherapy have also been investigated in microfluidic systems, where some devices were utilized to manufacture, harvest, and sequence MAbs [244][245][246][247]. The anti-ErbB-2 MAb trastuzumab has been prepared with radioactive 89 Zr for PET scanning in a microfluidic device, and the localization of trastuzumab to distal tumors was modeled in a similar device [248,249].…”

Multivariate Data Analysis Methodology to Solve Data Challenges Related to Scale‐Up Model Validation and Missing Data on a Micro‐Bioreactor System

“…Therefore, the variety, veracity, velocity, variability, and potential value within these biomanufacturing data sets warrants the classification of “Big Data.” Data analytics and the appropriate management of these small and large complex data sets represent a major challenge for the majority of industries including biopharmaceuticals. The increase in sensor technology, combined with recent advancements in parallelization and development of disposable bioreactors for scale‐down purposes has resulted in the production of a wide variety of complex data sets to interpret. The sheer volume and variety of the available data remains a significant unmet challenge within biopharmaceutical manufacturing.…”