Experiment and simulation of an aqueous two-phase extraction process for the purification of a monoclonal antibody

Mündges, J.; Zierow, Jessica; Zeiner, Tim

doi:10.1016/j.cep.2015.04.013

Cited by 15 publications

(8 citation statements)

References 29 publications

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“…By loading the top PEG rich phase into a cation exchange chromatography column, it was possible to bypass a conditioning step, while also reducing 10‐fold the amount of IgG aggregates in the end of the process [72]. Other research groups, lead by J. Hubbuch and T. Zeiner have also contributed to this field of research, using high‐throughput approaches [12, 82] and simulation and validation of continuous multi‐stage purification strategies, coupled with washing and back extraction steps [55, 93].…”

Section: Opportunities – Atps Partitioning In Relation To Other Modmentioning

confidence: 99%

Partitioning in aqueous two‐phase systems: Analysis of strengths, weaknesses, opportunities and threats

Soares

Azevedo

Alstine

et al. 2015

Biotechnology Journal

128

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For half a century aqueous two-phase systems (ATPSs) have been applied for the extraction and purification of biomolecules. In spite of their simplicity, selectivity, and relatively low cost they have not been significantly employed for industrial scale bioprocessing. Recently their ability to be readily scaled and interface easily in single-use, flexible biomanufacturing has led to industrial re-evaluation of ATPSs. The purpose of this review is to perform a SWOT analysis that includes a discussion of: (i) strengths of ATPS partitioning as an effective and simple platform for biomolecule purification; (ii) weaknesses of ATPS partitioning in regard to intrinsic problems and possible solutions; (iii) opportunities related to biotechnological challenges that ATPS partitioning may solve; and (iv) threats related to alternative techniques that may compete with ATPS in performance, economic benefits, scale up and reliability. This approach provides insight into the current status of ATPS as a bioprocessing technique and it can be concluded that most of the perceived weakness towards industrial implementation have now been largely overcome, thus paving the way for opportunities in fermentation feed clarification, integration in multi-stage operations and in single-step purification processes.

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Section: Opportunities – Atps Partitioning In Relation To Other Modmentioning

confidence: 99%

Partitioning in aqueous two‐phase systems: Analysis of strengths, weaknesses, opportunities and threats

Soares

Azevedo

Alstine

et al. 2015

Biotechnology Journal

128

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“…Although PEG is biodegradable and non‐toxic, phosphate disposal is problematic. These bottlenecks may however be minimized if the recycling of both PEG and phosphate is considered . In general, the strategy here proposed and followed showed ATPS to be a potential, industrially suited, primary recovery operation for human IgG.…”

Section: Discussionmentioning

confidence: 91%

“…In our case, the recovery was increased with top phase recirculation reaching a 78%, but decreasing purity to a 28%. As reported in other similar extraction works, in a multi‐stage equilibrium extraction, an increase in the number of stages allows an increase in purity, whether they are extraction, back extraction or washing steps . High recovery yields, although desirable, is not encouraged during the first stages since this issue is meant to be solved with further purification steps .…”

Section: Discussionmentioning

confidence: 96%

An integrated practical implementation of continuous aqueous two‐phase systems for the recovery of human IgG: From the microdevice to a multistage bench‐scale mixer‐settler device

Espitia-Saloma

Vázquez-Villegas

Rito‐Palomares

et al. 2016

Biotechnology Journal

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Aqueous two-phase systems (ATPS) are a liquid-liquid extraction technology with clear process benefits; however, its lack of industrial embracement is still a challenge to overcome. Antibodies are a potential product to be recovered by ATPS in a commercial context. The objective of this work is to present a more integral approach of the different isolated strategies that have arisen in order to enable a practical, generic implementation of ATPS, using human immunoglobulin G (IgG) as experimental model. A microfluidic device is used for ATPS parameters preselection for product recovery. ATPS were continuously operated in a mixer-settler device in one stage, multistage and multistage with recirculation configuration. Single-stage pure IgG extraction with a polyethylene glycol (PEG) 3350-phophates ATPS within continuous operation allowed a 65% recovery. Further implementation of a multistage platform promoted a higher particle partitioning reaching a 90% recovery. The processing of IgG from a cell supernatant culture harvest in a multistage system with top phase recirculation resulted in 78% IgG recovery in bottom phase. This work conjugates three not widely spread methodologies for ATPS: microfluidics, continuous and multistage operation.

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“…In the same line, Ziljstra et al [173,174] coupled triazine dye mimetic green to PEG in order to selectively recover IgG from hybridoma cells grown in the dextran-rich bottom phase. More recently, in 2015, Muendges et al [175,176] published two studies on the extraction and purification of IgG from CHO cell lines through a single-stage and by using a multi-stage aqueous two-phase extraction. In the single-stage approach, the authors [176] were able to extract IgG1 from a CHO cell culture supernatant with a system composed of PEG 2000 and sodium phosphate at pH 6 with an yield higher than 90% and purification factor up to 3.1.…”

Section: Aqueous Two-phase Systems (Atps)mentioning

confidence: 99%

Monoclonal Antibodies - Addressing the Challenges on the Manufacturing Processing of an Advanced Class of Therapeutic Agents

Capela¹,

Aires‐Barros²,

Freire³

et al. 2017

Frontiers in Clinical Drug Research: Anti-Infectives Volume 4

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Monoclonal antibodies (mAbs) were firstly described by Köhler and Milstein in 1975, and their potential as powerful therapeutic agents recognized in the following years. Currently, the US Food and Drug Administration (FDA) has already approved 56 monoclonal antibodies for the treatment of several diseases, including cancer (e.g. breast cancer, leukemia and prostate cancer), auto-immune disorders (rheumatoid arthritis and Crohn's disease), asthma, and cardiovascular and infectious diseases. Despite their advantages and therapeutic potential, the cost of manufacturing these biopharmaceuticals with high quality and purity level is still extremely high due to the absence of current cost-effective extraction/purification methods, and which has also impaired their widespread application as recurrent therapeutic agents. The upstream processing of mAbs has gone through several improvements in recent years, by using alternative expression systems or by optimizing the medium formulations and feeding strategies. On the contrary, the downstream processing is considered the bottleneck in the manufacturing of mAbs for therapeutic purposes at reliable costs, representing up to 80% of their total production costs-which is a frontier in clinical drug research. In the past years, several chromatographic and non-chromatographic alternatives have been explored for this purpose, resulting in the development of efficient platforms for the purification of mAbs, that are overviewed and discussed in this chapter. In summary, this chapter provides a vision on the current state of the art of the biopharmaceuticals market, on the production and use of mAbs as valuable therapeutic agents, including their use for the treatment of infectious diseases, while summarizing the mAbs-based products already approved by regulatory agencies. New insights concerning the development of new and alternative platforms or the extraction

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Experiment and simulation of an aqueous two-phase extraction process for the purification of a monoclonal antibody

Cited by 15 publications

References 29 publications

Partitioning in aqueous two‐phase systems: Analysis of strengths, weaknesses, opportunities and threats

Partitioning in aqueous two‐phase systems: Analysis of strengths, weaknesses, opportunities and threats

An integrated practical implementation of continuous aqueous two‐phase systems for the recovery of human IgG: From the microdevice to a multistage bench‐scale mixer‐settler device

Monoclonal Antibodies - Addressing the Challenges on the Manufacturing Processing of an Advanced Class of Therapeutic Agents

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