At-line process analytical technology (PAT) for more efficient scale up of biopharmaceutical microfiltration unit operations

Watson, Douglas S.; Kerchner, Kristi R.; Gant, Sean S.; Pedersen, Joseph W.; Hamburger, James B.; Ortigosa, Allison D.; Potgieter, Thomas I.

doi:10.1002/btpr.2193

Cited by 6 publications

(3 citation statements)

References 30 publications

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“…On‐line process analytical technology (PAT) would also need to be incorporated as part of any commercial continuous process. Watson et al recently showed how at line PAT could be incorporated in the scale up of a microfiltration unit; a similar approach could potentially be used to incorporate PAT in a CCTC system.…”

Section: Discussionmentioning

confidence: 99%

Performance optimization of continuous countercurrent tangential chromatography for antibody capture

Dutta

Tan

Zydney

et al. 2016

Biotechnology Progress

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Recent studies have demonstrated that continuous countercurrent tangential chromatography (CCTC) can effectively purify monoclonal antibodies from clarified cell culture fluid. CCTC has the potential to overcome many of the limitations of conventional packed bed protein A chromatography. This paper explores the optimization of CCTC in terms of product yield, impurity removal, overall productivity, and buffer usage. Modeling was based on data from bench-scale process development and CCTC experiments for protein A capture of two clarified Chinese Hamster Ovary cell culture feedstocks containing monoclonal antibodies provided by industrial partners. The impact of resin binding capacity and kinetics, as well as staging strategy and buffer recycling, was assessed. It was found that optimal staging in the binding step provides better yield and increases overall system productivity by 8-16%. Utilization of higher number of stages in the wash and elution steps can lead to significant decreases in buffer usage (∼40% reduction) as well as increased removal of impurities (∼2 log greater removal). Further reductions in buffer usage can be obtained by recycling of buffer in the wash and regeneration steps (∼35%). Preliminary results with smaller particle size resins show that the productivity of the CCTC system can be increased by 2.5-fold up to 190 g of mAb/L of resin/hr due to the reduction in mass transfer limitations in the binding step. These results provide a solid framework for designing and optimizing CCTC technology for capture applications. © 2016 American Institute of Chemical Engineers Biotechnol. Prog., 32:430-439, 2016.

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Section: Discussionmentioning

confidence: 99%

Performance optimization of continuous countercurrent tangential chromatography for antibody capture

Dutta

Tan

Zydney

et al. 2016

Biotechnology Progress

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“…Small production skids, disposable units, variable raw materials, and lack of real-time online measurement devices are challenges for PAT integration, but new, sophisticated instruments show promising improvements [82] . Product attribute control is now possible within the bioreactor, for example with MAST (Modular Automated Sampling Technology), and advances in Raman spectroscopy, at-line surface plasmon resonance (SPR), SU sensors, chemometric methods, and ultra high performance liquid chromatography (UPLC) at line monitoring are helping manufacturers overcome hurdles in DSP PAT integration [83] , [83] , [84] , [85] , [86] , [87] , [88] .…”

Section: Summary and Outlooksmentioning

confidence: 99%

A Single-use Strategy to Enable Manufacturing of Affordable Biologics

Jacquemart

Vandersluis

Zhao

et al. 2016

Computational and Structural Biotechnology Journal

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The current processing paradigm of large manufacturing facilities dedicated to single product production is no longer an effective approach for best manufacturing practices. Increasing competition for new indications and the launch of biosimilars for the monoclonal antibody market have put pressure on manufacturers to produce at lower cost. Single-use technologies and continuous upstream processes have proven to be cost-efficient options to increase biomass production but as of today the adoption has been only minimal for the purification operations, partly due to concerns related to cost and scale-up. This review summarizes how a single-use holistic process and facility strategy can overcome scale limitations and enable cost-efficient manufacturing to support the growing demand for affordable biologics. Technologies enabling high productivity, right-sized, small footprint, continuous, and automated upstream and downstream operations are evaluated in order to propose a concept for the flexible facility of the future.

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“…It was used to measure both protein concentration and purity across the entire process, including excluded chromatographic fractions. The method utilized a 2.1 × 150 mm column (Waters Acquity BEH Shield C18 1.7 µm, Catalog #186003376) and a gradient of 27%–33% acetonitrile over 28 min employing a water/acetonitrile/trifluoroacetic acid gradients at a flow rate of 0.2 mL min −1 on a Waters H‐Class UPLC instrument (Watson et al, 2016). Additionally, this same method was used to measure purity at release, further enabling an integrated control strategy across the entire process assuring the purity target was met at the final drug substance (DS).…”

Section: Introduction Key Elements and Themesmentioning

confidence: 99%

Insulin purification—Innovation continuum via synthesis of fundamentals, technology, and modeling

Roush

Iammarino

Chmielowski

et al. 2023

Biotech & Bioengineering

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Advancement in all disciplines (art, science, education, and engineering) requires a careful balance of disruption and advancement of classical techniques. Often technologies are created with a limited understanding of fundamental principles and are prematurely abandoned. Over time, knowledge improves, new opportunities are identified, and technology is reassessed in a different light leading to a renaissance. Recovery of biological products is currently experiencing such a renaissance. Crystallization is one example of an elegant and ancient technology that has been applied in many fields and was employed to purify insulins from naturally occurring sources. Crystallization can also be utilized to determine protein structures. However, a multitude of parameters can impact protein crystallization and the “hit rate” for identifying protein crystals is relatively low, so much so that the development of a crystallization process is often viewed as a combination of art and science even today. Supplying the worldwide requirement for insulin (and associated variants) requires significant advances in process intensification to support scale of production and to minimize the overall cost to enable broader access. Expanding beyond insulin, the increasing complexity and diversity of biologics agents challenge the current purification methodologies. To harness the full potential of biologics, there is a need to fully explore a broader range of purification technologies, including nonchromatographic approaches. This impetus requires one to challenge and revisit the classical techniques including crystallization, chromatography, and filtration from a different vantage point and with a new set of tools, including molecular modeling. Fortunately, computational biophysics tools now exist to provide insights into mechanisms of protein/ligand interactions and molecular assembly processes (including crystallization) that can be used to support de novo process development. For example, specific regions or motifs of insulins and ligands can be identified and used as targets to support crystallization or purification development. Although the modeling tools have been developed and validated for insulin systems, the same tools can be applied to more complex modalities and to other areas including formulation, where the issue of aggregation and concentration‐dependent oligomerization could be mechanistically modeled. This paper will illustrate a case study juxtaposing historical approaches to insulin downstream processes to a recent production process highlighting the application and evolution of technologies. Insulin production from Escherichia coli via inclusion bodies is an elegant example since it incorporates virtually all the unit operations associated with protein production—recovery of cells, lysis, solubilization, refolding, purification, and crystallization. The case study will include an example of an innovative application of existing membrane technology to combine three‐unit operations into one, significantly reducing solids handling and buffer consumption. Ironically, a new separations technology was developed over the course of the case study that could further simplify and intensify the downstream process, emphasizing and highlighting the ever‐accelerating pace of innovation in downstream processing. Molecular biophysics modeling was also employed to enhance the mechanistic understanding of the crystallization and purification processes.

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At-line process analytical technology (PAT) for more efficient scale up of biopharmaceutical microfiltration unit operations

Cited by 6 publications

References 30 publications

Performance optimization of continuous countercurrent tangential chromatography for antibody capture

Performance optimization of continuous countercurrent tangential chromatography for antibody capture

A Single-use Strategy to Enable Manufacturing of Affordable Biologics

Insulin purification—Innovation continuum via synthesis of fundamentals, technology, and modeling

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