Supermacroporous polymer‐based cryogel bioreactor for monoclonal antibody production in continuous culture using hybridoma cells

Abstract

Cryogel matrices composed of different polymeric blends were synthesized, yielding a unique combination of hydrophilicity and hydrophobicity with the presence or absence of charged surface. Four such cryogel matrices composed of polyacrylamide-chitosan (PAAC), poly(N-isopropylacrylamide)-chitosan, polyacrylonitrile (PAN), and poly(N-isopropylacrylamide) were tested for growth of different hybridoma cell lines and production of antibody in static culture. All the matrices were capable for the adherence of hybridoma cell lines 6A4D7, B7B10, and H9E10 to the polymeric surfaces as well as for the efficient monoclonal antibody (mAb) production. PAAC proved to be relatively better in terms of both mAb production and cell growth. Further, PAAC cryogel was designed into three different formats, monolith, disks, and beads, and used as packing material for packed-bed bioreactor. Long-term cultivation of 6A4D7 cell line on PAAC cryogel scaffold in all the three formats could be successfully done for a period of 6 weeks under static conditions. Continuous packed-bed bioreactor was setup using 6A4D7 hybridoma cell line in the three reactor formats. The reactors ran continuously for a period of 60 days during which mAb production and metabolism of cells in the bioreactors were monitored periodically. The monolith bioreactor performed most efficiently over a period of 60 days and produced a total of 57.5 mg of antibody in the first 30 days (in 500 mL) with a highest concentration of 115 μg mL(-1) , which is fourfold higher than t-flask culture. The results demonstrate that appropriate chemistry and geometry of the bioreactor matrix for cell growth and immobilization can enhance the reactor productivity.

Metabolic responses and pathway changes of mammalian cells under different culture conditions with media supplementations

“…A significant amount of work has been done in cell culture technology over the last few decades for improving mAb productivity with maintaining quality attributes while reducing manufacturing cost and time to market . To improve the mAb titer, cell line engineering, cell culture operation conditions, and media optimization have been studied. Another idea of improving mAb productivity can be obtained by increasing understanding of cell organisms by applying metabolic flux analysis.…”

Comparative Study of Various Delivery Methods for the Supply of Alpha-Ketoglutarate to the Neural Cells for Tissue Engineering

“…Every third day, the media were collected in triplicates and assayed for ammonia by indophenol method [21]. In brief, 1 g of phenol and 5 mg sodium nitroprusside were dissolved in 100 mL deionized water to get solution I.…”

Evaluation of the effectiveness of microparticle-embedded cryogel system in removal of 17β-estradiol from aqueous solution

“…Due to the high porosity, high permeability, soft, flexibility, and high flow velocity cryogels are widely used as new chromatographic materials for separation and purification of biomolecules, such as proteins, plasmid DNA, and viruses, from crude feedstocks [15][16][17] and also different applications as immobilization matrices in biotechnology [18][19][20][21], as scaffolds in tissue engineering [22][23][24][25][26], and as drug-delivery carriers in pharmaceutical fields [27]. Cryogels permit the free passage of microparticles, nanoparticles, or bioparticles without blockage because of the pore size within the range of 10-100 μm.…”