State‐of‐the‐art in downstream processing of monoclonal antibodies: Process trends in design and validation

Abstract

Monoclonal antibodies (mAbs) are the most important family of biopharmaceutical compounds in terms of market share. At present, 30 mAbs have been approved and are now commercialized for therapeutic purposes. mAbs are typically produced by mammalian cell culture in bioreactors that range in scale of 1-20 m(3) . Regardless of scale, from laboratory to commercial settings, the recovery and purification of mAbs present important challenges. Depending on the scale, the particular product, and the type of production process (bioreactor operation, process time, complexity of the culture media, cell density, etc.), many possible downstream configurations are possible and have been used. In this contribution, we review each type of unit operation that forms a downstream train for mAb production. We provide information regarding typical operation settings and critical variables for centrifugation, ultrafiltration, affinity chromatography, ion exchange chromatography, and viral removal operations. In addition, we discuss some important considerations required for the formulation of drugs based on mAbs.

Comprehensive glycosylation profiling of IgG and IgG-fusion proteins by top-down MS with multiple fragmentation techniques

“…The mAb was expressed in stably transfected NS0 cells and purified using a standard multi-step purification process [16]. It is a fully human monoclonal antibody (IgG1).…”

“…The product quality of the mAb was confirmed by non-reducing gel electrophoresis (96.0% purity) and high performance size exclusion chromatography (99.7% purity). The fusion protein was expressed in stably transfected Chinese hamster ovary cells (CHO) and purified using a standard multi-step purification process [16]. The product quality of the IgG fusion protein was confirmed by non-reducing gel electrophoresis (99.0% purity) and high performance size exclusion chromatography (99.8% purity).…”

Model-Based Comparison of Antibody Dimerization in Continuous and Batch-Wise Downstream Processing

“…The use of monoclonal antibodies as biopharmaceuticals is becoming more popular [1,2], and thus antibody dimerization during preparation and the subsequent removal of aggregates are becoming increasingly studied areas. When modeling the purification of antibodies from dimers, the main focus has been on the removal of dimers produced during the upstream processing.…”

“…Monoclonal antibodies are usually produced in a bioreactor, and a series of separation steps is subsequently used to isolate and purify the product [2][3][4]. This downstream processing normally consists of chromatographic purification steps that are run in batch mode, with storage tanks between each unit operation.…”

IMAC capture of recombinant protein from unclarified mammalian cell feed streams

“…Typically, solid–liquid separation consists of centrifugation and filtration. These techniques can account for up to 25% COG/g when factors such as capital cost and energy consumption are included (Marichal‐Gallardo and Álvarez, 2012). Primary processing of high cell density feed streams requires particular consideration to avoid cell stresses and shearing that could result in protein aggregation and cell lysis (Hutchinson et al, 2006; Kiese et al, 2008).…”