A fast and simple approach to optimize the unit operation high pressure homogenization - a case study for a soluble therapeutic protein in<i>E. coli</i>

Pekarsky, Alexander; Spadiut, Oliver; Rajamanickam, Vignesh; Wurm, David Johannes

doi:10.1080/10826068.2018.1536988

Cited by 11 publications

(4 citation statements)

References 22 publications

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“…Primary recovery in E. coli processes usually requires cell disruption by high‐pressure homogenization to make the product available for purification. Homogenization is a critical unit operation that may have a large impact on further downstream steps, for example by introducing variance in pH or product concentration, and therefore requires optimization 23–25 . The major advantage of extracellular protein production is that primary recovery is simplified by omitting this cell disruption step.…”

Section: Resultsmentioning

confidence: 99%

Economic and ecological benefits of a leaky E. coli strain for downstream processing: a case study for staphylococcal protein A

Kastenhofer

Cataldo

Ebner

et al. 2021

J of Chemical Tech & Biotech

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BACKGROUND Downstream processing of soluble recombinant proteins from Escherichia coli is complicated by the need to access the intracellular product by cell disruption and to separate the target protein from impurities, particularly host cell protein, DNA, endotoxins and lipids. We previously demonstrated the ability of the E. coli X‐press strain to leak high amounts of product to the culture medium without sacrificing viability. In this case study, we assessed the economic and ecological benefit of this strain for downstream processing in direct comparison to the industrial standard E. coli BL21(DE3). Staphylococcal Protein A was used as a model protein. We performed recombinant protein production, primary recovery and capture by anion exchange chromatography at laboratory scale, and used the data obtained for estimating costs and resource consumption by economic modeling. RESULTS After primary recovery, the X‐press process resulted in a 1.5‐fold higher product purity, a 150‐fold lower DNA, 3.5‐fold lower endotoxin and 3.4‐fold lower lipid load compared to BL21(DE3). Consequently, anion exchanger binding capacity was increased 2.7‐fold and purity and concentration of the eluate also was increased. Extracellular protein production with X‐press resulted in a 25% reduction of costs and a 36% reduction of both water usage and water‐related CO2 emissions compared to intracellular production with BL21(DE3). CONCLUSIONS This case study performed with stapyhlococcal Protein A demonstrated the potential of E. coli X‐press to reduce costs for downstream processing and improve the environmental footprint by simplified primary recovery, lower impurity load and consequently higher chromatographic efficiency. © 2021 The Authors. Journal of Chemical Technology & Biotechnology published by John Wiley & Sons Ltd on behalf of Society of Chemical Industry.

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Section: Resultsmentioning

confidence: 99%

Economic and ecological benefits of a leaky E. coli strain for downstream processing: a case study for staphylococcal protein A

Kastenhofer

Cataldo

Ebner

et al. 2021

J of Chemical Tech & Biotech

Self Cite

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“…The use of DoE has also been established to increase the performance of downstream process development. In one study, high-pressure homogenization was used to screen critical process parameters (CPPs) using DoE to enhance product titer and achieve adequate product quality, based on predefined critical quality attributes (CQAs) (Pekarsky et al, 2019). A process for the purification of scFv using mixed-mode chromatography was developed using DoE, and it was found that the optimized conditions enabled binding of the scFv to Capto Adhere TM below its theoretical pI, with the majority of HCPs in the flow-through (Sakhnini et al, 2019).…”

Section: Design Of Experiments (Doe) Approachmentioning

confidence: 99%

Recent Developments in Bioprocessing of Recombinant Proteins: Expression Hosts and Process Development

Tripathi

Shrivastava

2019

Front. Bioeng. Biotechnol.

369

243

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Infectious diseases, along with cancers, are among the main causes of death among humans worldwide. The production of therapeutic proteins for treating diseases at large scale for millions of individuals is one of the essential needs of mankind. Recent progress in the area of recombinant DNA technologies has paved the way to producing recombinant proteins that can be used as therapeutics, vaccines, and diagnostic reagents. Recombinant proteins for these applications are mainly produced using prokaryotic and eukaryotic expression host systems such as mammalian cells, bacteria, yeast, insect cells, and transgenic plants at laboratory scale as well as in large-scale settings. The development of efficient bioprocessing strategies is crucial for industrial production of recombinant proteins of therapeutic and prophylactic importance. Recently, advances have been made in the various areas of bioprocessing and are being utilized to develop effective processes for producing recombinant proteins. These include the use of high-throughput devices for effective bioprocess optimization and of disposable systems, continuous upstream processing, continuous chromatography, integrated continuous bioprocessing, Quality by Design, and process analytical technologies to achieve quality product with higher yield. This review summarizes recent developments in the bioprocessing of recombinant proteins, including in various expression systems, bioprocess development, and the upstream and downstream processing of recombinant proteins.

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“…Many studies also analyze process and product specific nuances, exploring avenues for improved efficiency, quality, and control. These include investigating the interplay between upstream and downstream unit operations on inclusion body quality and yield (Eggenreich et al, 2020;Slouka et al, 2018), characterizing the impact of processinduced chemical modifications to monoclonal antibodies (Hutterer et al, 2013), improved cell disruption process monitoring strategies (Eggenreich et al, 2017), and strategies for process optimization (Dopp and Reuel, 2018;Metzger et al, 2020;Pekarsky et al, 2019).…”

Section: Biopharmaceutical and Biotechnology Applicationsmentioning

confidence: 99%

High Pressure Homogenization – An Update on its Usage and Understanding

Inguva,

Grasselli,

Heng

2023

Preprint

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Despite being a technology of several decades, high pressure homogenization (HPH) remains widely used in food and pharmaceutical industries, often as an essential unit operation in liquid product processing. Continual advances in the technology are made on multiple fronts, on equipment innovations by the manufacturers, new applications by users, and advances in process understanding by multidisciplinary scientists alongside subject matter experts amongst industry practitioners. While HPH is comparatively simple conceptually, the homogenization process involves complex engineering physics which is influenced by the varied processing conditions and highly diverse inputs with each use-case requiring its own treatment. The successful application of a HPH process indubitably requires practitioners to draw upon insights from multiple domains and the optimization for each case. Thus, this timely review aims to outline the more recent trends and advancements in HPH process understanding and novel applications involving HPH from both academic and industrial perspectives.

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A fast and simple approach to optimize the unit operation high pressure homogenization - a case study for a soluble therapeutic protein inE. coli

Abstract: View related articles View Crossmark data Citing articles: 2 View citing articles A fast and simple approach to optimize the unit operation high pressure homogenization-a case study for a soluble therapeutic protein in E. coli

Cited by 11 publications

References 22 publications

Economic and ecological benefits of a leaky E. coli strain for downstream processing: a case study for staphylococcal protein A

Economic and ecological benefits of a leaky E. coli strain for downstream processing: a case study for staphylococcal protein A

Recent Developments in Bioprocessing of Recombinant Proteins: Expression Hosts and Process Development

High Pressure Homogenization – An Update on its Usage and Understanding

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