Application of an aqueous two-phase systems strategy for the potential recovery of a recombinant protein from alfalfa (Medicago sativa)

Ibarra‐Herrera, Celeste C.; Aguilar, Oscar; Rito‐Palomares, Marco

doi:10.1016/j.seppur.2010.11.025

Cited by 53 publications

(33 citation statements)

References 27 publications

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“…On the other hand, increasing corn solids and lysozyme loading did not change the purification factor significantly until 10× loading for germ [7]. Partition coefficients of native protein, which are extracted from corn germ [7], tobacco [22,23], and soybean [3,43], are generally significantly less than 1, even with NaCl addition, while native protein from alfalfa [44,45] and corn endosperm [7] partitioned more to top phase with NaCl addition.…”

Section: Effect Of Atp Systems' Parametersmentioning

confidence: 99%

“…It has been demonstrated for different plant hosts, including corn [7], tobacco [22,23], alfalfa [44,45], and soybean [43,48], that an ATPP can be chosen and optimized for recovery of hydrophobic proteins in top phase from plant tissue/seed extracts, while at 20-40 % native protein from high-protein hosts, such as tobacco, alfalfa, soybean, and corn germ, forms precipitates on the interface between two phases. Furthermore, in PEG-salt ATP systems, partition coefficients of native protein, which are extracted from corn germ [7], tobacco [22], and soybean [43,48], are generally less than 1.…”

Section: Protein Propertiesmentioning

confidence: 99%

“…Furthermore, in PEG-salt ATP systems, partition coefficients of native protein, which are extracted from corn germ [7], tobacco [22], and soybean [43,48], are generally less than 1. But corn endosperm protein and alfalfa protein partitioned evenly into two phases or more to top phase than bottom phase [44,45] with NaCl addition. In addition, extraction can be advantageously integrated with the partitioning step.…”

Section: Protein Propertiesmentioning

confidence: 99%

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Recovery of Recombinant Proteins from Plants Using Aqueous Two-Phase Partitioning Systems: An Outline

2014

Methods in Molecular Biology

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Transgenic plants producing recombinant proteins are a potential low-cost substitute for complex bioreactors. However, the development of new robust downstream processes to improve protein recovery and isolation from plant feedstock is critical to promote this new technique because downstream processing costs typically contribute to more than 80 % of the total cost. Polishing separation platforms for protein purification, such as chromatography and membrane filtration, have been well established, while little attention has been allocated to initial concentration and separation procedures. In this chapter, application of aqueous two-phase partitioning (ATPP), as an attractive alternative to traditional processes for recovering and isolating target proteins from plant green tissues or seeds, as well as the main advances reported in literature concerning ATPP for the isolation and purification of proteins from plant feedstock are reviewed. The potential application of ATPP as an integrated extraction and isolation step and isolation step after extraction or protein characterization method is discussed separately. The connection of ATPP with traditional protein separation processes is discussed. The separation mechanisms of ATPP are explained based on surface properties of proteins and polymer systems. Finally, the future trends in applying ATPP for protein separation are discussed.

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Section: Effect Of Atp Systems' Parametersmentioning

confidence: 99%

Section: Protein Propertiesmentioning

confidence: 99%

Section: Protein Propertiesmentioning

confidence: 99%

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Recovery of Recombinant Proteins from Plants Using Aqueous Two-Phase Partitioning Systems: An Outline

2014

Methods in Molecular Biology

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“…The slurry is stirred for 1.0 h and then centrifuged at 12,000 × g for 10 min at room temperature. The supernatant is then fi ltered using 0.45 μm syringe fi lter [ 13 ].…”

Section: Green Tissue Protein Extractsmentioning

confidence: 99%

“…Lane 4 is top-phase sample corresponding to rhG-CSF (reproduced from ref. [ 13 ] with permission from Elsevier)…”

Section: Aqueous Two-phase Partitioningmentioning

confidence: 99%

Aqueous Two-Phase System Strategies for the Recovery of Proteins from Plants

Aguilar

Rito‐Palomares

2014

Methods in Molecular Biology

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The increasing interest of the biopharmaceutical industry to exploit plants as a commercially viable production system is demanding the development of new strategies to maximize product recovery. Aqueous two-phase systems (ATPSs) are a primary recovery technique that has shown great potential for the efficient extraction and purification of biological products. The evaluation of different system parameters upon the partitioning behavior can provide the conditions that favor the concentration of contaminants and the desired target protein in opposite phases. The protocols described provide the basic strategy to explore the use of ATPS for the isolation and partial purification of native and recombinant proteins expressed in plants.

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The Impact of Six Critical Impurities on Recombinant Protein Recovery and Purification from Plant Hosts

Dixon

Wilken

Woodard³

et al. 2018

Molecular Pharming

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Application of an aqueous two-phase systems strategy for the potential recovery of a recombinant protein from alfalfa (Medicago sativa)

Cited by 53 publications

References 27 publications

Recovery of Recombinant Proteins from Plants Using Aqueous Two-Phase Partitioning Systems: An Outline

Recovery of Recombinant Proteins from Plants Using Aqueous Two-Phase Partitioning Systems: An Outline

Aqueous Two-Phase System Strategies for the Recovery of Proteins from Plants

The Impact of Six Critical Impurities on Recombinant Protein Recovery and Purification from Plant Hosts

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