Optimizing refolding and recovery of active recombinant Bacillus halodurans xylanase in polymer–salt aqueous two-phase system using surface response analysis

Rahimpour, Farshad; Mamo, Gashaw; Feyzi, Farzaneh; Maghsoudi, Saeid; Hatti‐Kaul, Rajni

doi:10.1016/j.chroma.2006.11.053

Cited by 38 publications

(40 citation statements)

References 23 publications

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“…PEG interacts with the hydrophobic side chains exposed upon unfolding at high temperatures [46]. PEG has been successfully used for refolding of interferon [47], xylanase [48], and insulin-like growth factor (IGF-1) [49] from inclusion bodies.…”

Section: Polyethylene Glycol (Peg)mentioning

confidence: 99%

Retracted: Chemical Assistance in Refolding of Bacterial Inclusion Bodies

Alibolandi¹,

Mirzahosei²

2011

American J. of Biochemistry and Molecular Biology

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Escherichia coli is one of the most widely used hosts for the production of recombinant proteins but insoluble expression of heterologous proteins is a major bottleneck in production of recombinant proteins in E. coli. In vitro refolding of inclusion body into proteins with native conformations is a solution for this problem but there is a need for optimization of condition for each protein specifically. Several approaches have been described for in vitro refolding; most of them involve the use of additives for assisting correct folding. Cosolutes play a major role in refolding process and can be classified according to their function as aggregation suppressors and folding enhancers. This paper presents a review of additives that are used in refolding process of insoluble recombinant proteins in small scale and industrial processes.

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Section: Polyethylene Glycol (Peg)mentioning

confidence: 99%

Retracted: Chemical Assistance in Refolding of Bacterial Inclusion Bodies

Alibolandi¹,

Mirzahosei²

2011

American J. of Biochemistry and Molecular Biology

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“…However, when the experimental region of interest is cuboidal rather than rotatable/ spherical or when it is preferable to maintain low and high factor levels, and still carry out a RSM design, the CCF or face-centered cube is a feasible alternative involving α = ± 1 [9,13], which is commonly used for the optimization of PEG-salt ATPS [2,16,17,22,25,28,31,34,36,[49][50][51][52][53][54][55][56][57]. For instance, in the CCF design for three factors (e.g., x 1 , x 2 , and x 3 ) the star/axial points are located on the centers of the faces of a cube or hypercube (Fig.…”

Section: Accepted M Manuscriptmentioning

confidence: 99%

“…Moreover, FFD and fFD usually include 3-5 replicated center-point experiments which are carried out for an evaluation of the pure experimental error [9,13,[15][16][17][18]. These replicated experiments are called center-points because they are performed in the center of the experimental region located between the high and low levels, thus increasing the factor levels, at which the center level is denoted by 0 in a design matrix [13].…”

Section: Screening Of Significant Factorsmentioning

confidence: 99%

mentioning

confidence: 99%

“…In recent years, the use of RSM has gained importance in the evaluation of biotechnological processes and has become an innovative method in several research studies [17,28,29]. Furthermore, this approach is used as an effective statistical technique in order to determine the optimal operating conditions and significant independent factors or their interactions with the dependent output responses in multivariate complex systems, such as PEG-salt ATPS, thus enhancing the output responses for the optimization of these systems considering the partitioning of biomolecules, and consequently designing complex processes occurring in ATPS [2,11,17,19,[30][31][32]. Moreover, RSM is a useful and promising tool for establishing a statistical model to predict output responses or partitioning target parameters of ATPS by a correlation of the measured output response factors to the input factors, thus modelling a process, such as the separation and purification of biomolecules in ATPS [2,[32][33][34][35][36].…”

mentioning

confidence: 99%

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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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Disulfide–bridging PEGylation during refolding for the more efficient production of modified proteins

Ginn

Choi²,

Brocchini

2016

Biotechnology Journal

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Proteins that are modified by chemical conjugation require at least two separate purification processes. First the bulk protein is purified, and then after chemical conjugation, a second purification process is required to obtain the modified protein. In an effort to develop new enabling technologies to integrate bioprocessing and protein modification, we describe the use of disulfide-bridging conjugation to conduct PEGylation during protein refolding. Preliminary experiments using a PEG-mono-sulfone reagent with partially unfolded leptin and unfolded RNAse T1 indicated that the cysteine thiols underwent disulfide-bridging conjugation to give the PEGylated proteins. Interferon-β1b (IFN-β1b) was then expressed in E.coli as inclusion bodies and found to undergo disulfide bridging-conjugation during refolding. The PEG-IFN-β1b was isolated by ion-exchange chromatography and displayed in vitro biological activity. In the absence of the PEGylation reagent, IFN-β1b refolding was less efficient and yielded protein aggregates. No PEGylation was observed if the cysteines on IFN-β1b were first modified with iodoacetamide prior to refolding. Our results demonstrate that the simultaneous refolding and disulfide bridging PEGylation of proteins could be a useful strategy in the development of affordable modified protein therapeutics.

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Optimizing refolding and recovery of active recombinant Bacillus halodurans xylanase in polymer–salt aqueous two-phase system using surface response analysis

Cited by 38 publications

References 23 publications

Retracted: Chemical Assistance in Refolding of Bacterial Inclusion Bodies

Retracted: Chemical Assistance in Refolding of Bacterial Inclusion Bodies

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

Disulfide–bridging PEGylation during refolding for the more efficient production of modified proteins

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