Application of central composite design to the optimisation of aqueous two-phase extraction of human antibodies

Rosa, P.A.J.; Azevedo, Ana M.; Aires-Barros, M. Raquel

doi:10.1016/j.chroma.2006.11.075

Cited by 200 publications

(144 citation statements)

References 18 publications

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“…More than 95% of all impurities were removed, however, at the expense of a very low BE recovery yield (34%). The high NaCl concentrations determined for this BE system (7.4 and 6.5% in the BP and TP, respectively) together with the high volume ratio used may explain why such a low recovery yield was obtained for this step, as previous studies have shown that high NaCl concentration was one of the driving forces for the IgG extraction into the PEG-rich TP [4,11,15,19,26].…”

Section: Back-extraction Studiesmentioning

confidence: 82%

“…An Excellgene proprietary serum-free medium containing only one protein was used for production, and phenol red was added to the medium as a pH indicator. According to the ELISA determinations performed by Excellgene, the IgG titer was very low and was spiked with IgG from Gammanorm (pI of about 9 [4]) to final titers of 0.56 and 1.58 g/L. The CS 2 consisted of a PER.C6  CS containing IgG 1 (pI of about 8.5) and was provided by DSM Biologics (Groningen, The Netherlands).…”

Section: Reagentsmentioning

confidence: 99%

“…2013, 8,[352][353][354][355][356][357][358][359][360][361][362] www.biotecvisions.com downstream processing of mAbs includes clarification, concentration, selective purification steps, and virus clearance [3]. The selective purification steps usually comprise the mAbs adsorption to a protein A resin, followed by two further chromatography steps which remove host cell proteins (HCP), DNA, aggregates, any leached protein A and provide an adequate level of overall viral removal [3,4]. Although chromatographic separations have been the workhorse of most purification processes, several limitations have been pointed out, such as (i) batch operation, (ii) low capacity, (iii) complex scale-up, (iv) time-consuming and high pressure packing processing, (v) slow intraparticle diffusion, (vi) low chemical and proteolytic stability and consequent contamination of the final product, and (vii) the high cost of the resins [5][6][7][8][9].…”

mentioning

confidence: 99%

“…Aqueous two-phase system (ATPS) has shown to be considerably advantageous in terms of process economy and operation, especially when processing high titer cell culture supernatants [13]. This liquid-liquid extraction (LLE) technology has been successfully reported in recent times as a primary stage unit operation in the downstream processing of several biological products including therapeutic proteins, such as antibodies [4,10,11,[15][16][17][18][19][20][21][22][23][24]. Its technical feasibility at large scale has been, however, demonstrated for the downstream processing of just few biological products [25][26][27].…”

mentioning

confidence: 99%

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Continuous purification of antibodies from cell culture supernatant with aqueous two‐phase systems: From concept to process

Rosa

Azevedo

Sommerfeld

et al. 2013

Biotechnology Journal

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An aqueous two‐phase extraction (ATPE) process based on a PEG/phosphate system was developed for the capture of human immunoglobulin G and successfully applied to a Chinese hamster ovary and a PER.C6® cell supernatant. A continuous ATPE process incorporating three different steps (extraction, back‐extraction, and washing) was set up and validated in a pump mixer‐settler battery. Most of the higher molecular weight cell supernatant impurities were removed during the extraction step, while most of the lower molecular weight impurities were removed during the subsequent steps. A global recovery yield of 80% and a final protein purity of more than 99% were obtained for the IgG purification from a CHO cell supernatant, representing a 155‐fold reduction in the protein/IgG ratio. For the purification of IgG from a PER.C6® cell supernatant, a global recovery yield of 100%, and a host cell protein purity were attained, representing a 22‐fold reduction in the host cell protein/IgG ratio. These results, thus, open promising perspectives for the application of the developed ATPE process as a platform for the capture of antibodies. In fact, this new process has shown the ability to successfully recover and purify different antibodies from distinct cell culture supernatants. This technology can also overcome some of the limitations encountered using the typical chromatographic processes, besides inherent advantages of scalability, process integration, capability of continuous operation, and economic feasibility.

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Section: Back-extraction Studiesmentioning

confidence: 82%

Section: Reagentsmentioning

confidence: 99%

mentioning

confidence: 99%

mentioning

confidence: 99%

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Continuous purification of antibodies from cell culture supernatant with aqueous two‐phase systems: From concept to process

Rosa

Azevedo

Sommerfeld

et al. 2013

Biotechnology Journal

Self Cite

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“…(ANOVA) in order to calculate and evaluate the main effects of each factor and all their possible interactions on the output response variables [7,9,22].…”

Section: Screening Of Significant Factorsmentioning

confidence: 99%

Optimization of PEG–salt aqueous two-phase systems by design of experiments

Glyk

Solle

Scheper

et al. 2015

Chemometrics and Intelligent Laboratory Systems

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Since the mechanism governing the partitioning behavior of biomolecules, such as proteins and enzymes, in polyethylene glycol (PEG)-salt aqueous two-phase systems (ATPS) is complex and not easily predictable, many laborious experiments have to be performed for an optimization of these systems, causing increased overall cost. However, the multivariate statistical design of experiments (DoE) methodology is representing a promising and efficient optimization technique which can overcome the limitations of traditional optimization methods. Therefore, DoE has emerged as a powerful and efficient optimization tool for PEGsalt ATPS, since it is faster, more efficient and cost-effective, allowing a simultaneous and rigorous evaluation of process/system parameters. In the present review, different DoE process steps are represented to highlight the feasibility of this approach to operate as a promising and efficient optimization tool, thus facilitating the evaluation of the partitioning behavior, recovery and purification of different proteins and enzymes in PEG-salt ATPS. In this context, several experimental designs, such as factorial and response surface designs, have been discussed and evaluated by statistical regression analysis and analysis of variance (ANOVA), as well as various applications of PEG-salt ATPS using DoE have been outlined which may further promote the optimization of these systems.

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Ethylene Oxide Polymers: Synthesis, Modification, Topology, and Applications

Shen

Wang

2018

Encyclopedia of Polymer Science and Technology

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This article reviews the synthesis, modification, topology, and applications of ethylene oxide polymers (PEGs). The synthetic strategies presented involve the anionic ring‐opening polymerization, cationic ring‐opening polymerization, coordination polymerization, aluminum‐activated polymerization, and metal‐free polymerization. The modification concerns the m PEG derivatives, telechelic and heterotelechelic PEG. PEGs topologies discussed are focused on the block, star‐shaped, hyperbranched, and dendrimer‐like polymers. Based on the well‐defined modification and topology construction of PEGs, their applications in bioconjugation (or PEGylation) and pharmaceutical field, as well as in solid electrolyte, separation science, and support resins are presented. The article aims to help the readers to comprehensively understand the recent progress on PEGs and motivate further interest on these “evergreen” polymers.

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Application of central composite design to the optimisation of aqueous two-phase extraction of human antibodies

Cited by 200 publications

References 18 publications

Continuous purification of antibodies from cell culture supernatant with aqueous two‐phase systems: From concept to process

Continuous purification of antibodies from cell culture supernatant with aqueous two‐phase systems: From concept to process

Optimization of PEG–salt aqueous two-phase systems by design of experiments

Ethylene Oxide Polymers: Synthesis, Modification, Topology, and Applications

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