Biomedical Engineering and Bioengineering Commons^™

Design And Control Of Chromatography Step In An Integrated Column Sequence, Bernt Nilsson, Niklas Andersson, Anders Holmqvist, Anton Sellberg, Peter Tiainen, Arne Staby

Integrated Continuous Biomanufacturing II

Integration of the purification steps in downstream processing is one way to minimize the number of storage tanks and reduce hold-up time. The result is an integrated column sequence that performs straight through processing of the target protein, with minimal time from expression to formulation. This paper discusses the overall integration design and its expected performance, together with a more detailed analysis of the step behavior based on column simulations. The optimal design of the individual steps in the sequence is presented, both with robustness margins and local control system, as well as with and without local recycle.

An optimization …

Laboratory Scale Continuous Linear Purification As A Development Tool For Recombinant Blood Protein Processing, Using Chromatographic Resins And Membranes, Michael Hughson, Rimenys Carvalho, Thaynna Araujo Cruz, Leda Dos Reis Castilho

Integrated Continuous Biomanufacturing II

Continuous processing offers significant advantages for the processing of unstable recombinant products such as therapeutic plasma proteins. As such we have established a development platform to assess potential purification steps as part of a continuous linear process using a standard AKTA Explorer. Following a consistent format of IEX membrane to HIC membrane to affinity resin, we were able to rapidly investigate multiple purification pathways for a recombinant blood protein. Using membranes as the first two steps enables a 3-step purification process to be carried out in 3 to 4 hours, with a total turnaround of 5 to 6 hours including …

Enabling Technologies For Integrated / Continuous Downstream Processing Of Biologics, Jeff Salm, Marcus Fiadeiro, Raquel Orozco, Jill Kublbeck, Aaron Noyes, Jeff Horne, Daniel Lacasse, Ashley Sacramo, Suhani Gupta, John Coffman, Robert Fahrner

Integrated Continuous Biomanufacturing II

Pfizer Bioprocessing R&D is focused on developing enabling technologies that will reduce capital and operational expenses, decrease equipment scale, increase automation and utilize fewer FTEs.

To realize this vision, Purification Process Development has piloted new technologies and operational strategies that have enabled a fully integrated downstream process. Our current work has demonstrated a continuous process that includes tangential flow filtration harvest from a perfusion bioreactor, Protein A capture, inline viral inactivation/conditioning and AEX polishing. This process was fully automated and demonstrated at the 100 L scale. We have also shown feasibility of multi-day virus reduction filter operation and a continuous …

Spectral Deconvolution Of Chromatograms Without Offline Analytics, Matthias Rudt, Nina Brestrich, Florian Dismer, Jurgen Hubbuch

Integrated Continuous Biomanufacturing II

Continuous bioprocessing has shifted into the focus of the pharmaceutical industry in recent past. An important driving force is the expected higher productivity compared to operations in batch mode. To ensure constant quality attributes over a complete production cycle, reliable process analytical technologies (PAT) are central.

The deconvolution of spectral measurements in downstream processing has been introduced previously as a powerful PAT tool[1]. For spectral deconvolution, published research focused on partial least square (PLS) regression models. PLS models are able to deconvolute spectra even if the spectral differences of proteins are minute. They are widely used in chemometrics. However, PLS …

Toward Complete Continuity In Antibody Biomanufacture: Multi-Column Continuous Chromatography For Protein A Capture And Mixed Mode Hydroxyapatite Polishing, Anthony Grabski, Tom Van Oosbree, Beth Hammer, Alla Zilberman, Robert Mierendoff

Integrated Continuous Biomanufacturing II

Monoclonal antibodies (mAbs) are a predominant modality for a broad range of clinical indications including oncology and inflammatory diseases. Increasing manufacturing capacity and decreasing cost per purification campaign are critical factors for making antibody therapies more affordable. Cell culture mAb expression levels have steadily increased over the past ten years with titers of five grams per liter frequently achieved. Higher titers reduce production costs and allow processing of kilogram quantities for clinical trials from single use cell culture vessels. Drawbacks of increased titers include higher levels of aggregates, fragments, variants, and process impurities. These combined titer and impurity burdens further …