Monoliths in Bioprocess Technology

Abstract

Monolithic columns are a special type of chromatography column, which can be used for the purification of different biomolecules. They have become popular due to their high mass transfer properties and short purification times. Several articles have already discussed monolith manufacturing, as well as monolith characteristics. In contrast, this review focuses on the applied aspect of monoliths and discusses the most relevant biomolecules that can be successfully purified by them. We describe success stories for viruses, nucleic acids and proteins and compare them to conventional purification methods. Furthermore, the advantages of monolithic columns over particle-based resins, as well as the limitations of monoliths are discussed. With a compilation of commercially available monolithic columns, this review aims at serving as a 'yellow pages' for bioprocess engineers who face the challenge of purifying a certain biomolecule using monoliths.

Biopharmaceuticals from microorganisms: from production to purification

“…In addition, monolith does not have the packing step and tolerates the passage of air, reducing costs, and time with packing validation and repacking/replacing solid phase due to air interruption. Other significant advantages are easy scale-up due to flow independent of dynamic binding and compatibility with several organic, polymer-based, and inorganic media 69 . The disadvantage of higher buffer consumption can be decreased with the SMB set up, which can also be combined with single use technology.…”

“…Monoliths are widely applied to the recovery of proteins such as coagulation factor IX (ion exchange) 70 and IgG (affinity chromatography) 29 from a variety of cell culture including P. pastoris 71 and E. coli. 69 …”

An automated data‐driven DSP development approach for glycoproteins from yeast

“…In a previous study, we used monolithic columns for purification of three HRP isoenzymes differing in the number of N‐glycosylation sites with a two‐step flowthrough mode purification. Irrespective of the number of N‐glycosylation sites, the target recombinant protein was always found in the flowthrough fraction for monolithic columns, which was not the case using particle‐based resins . The SDS PAGE of fractions from the flowthrough mode purification of recombinant HRP from yeast, is shown in Supporting Information Fig.…”

“…We speculated that the glycans on the target protein surface shield the physicochemical properties of the biomolecule, allowing flowthrough mode purification. The major advantages of monolithic columns are high mass transfer properties, high binding capacity, customizability and scalability .…”

“…UV chromatograms have also been implemented for real‐time pooling decisions in DSP processes . Attributing to the fact that the target protein was always found in the flowthrough for monolithic columns , we used data driven approaches to calculate the total amount of impurities separated from the target protein before.…”