A metal‐chelate affinity reverse micellar system for protein extraction

Dong, Xiaoyan; Meng, Yao; Feng, Xudong; Sun, Yan

doi:10.1002/btpr.291

Cited by 16 publications

(16 citation statements)

References 45 publications

(61 reference statements)

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“…One milliliter of supernatant was carefully collected into fresh vials and lyophilized to obtain a solid pellet. In the organic phase, the orientations of hydrophobic and hydrophilic segments are reversed to form reversed (25) and release the contents. Addition of dichloromethane reverses the MNF and release the drug in the surrounding organic environment.…”

Section: Entrapment and Loading Efficiencymentioning

confidence: 99%

Optimization of Dexamethasone Mixed Nanomicellar Formulation

et al. 2014

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Abstract. The purpose of this study was to develop a clear aqueous mixed nanomicellar formulation (MNF) of dexamethasone utilizing both D-α-tocopherol polyethylene glycol-1000 succinate (Vit E TPGS) and . In this study, Vit E TPGS and Oc-40 are independent variables. Formulations were prepared following solvent evaporation method. A three level full-factorial design was applied to optimize the formulation based on entrapment efficiency, size, and polydispersity index (PDI). A specific blend of Vit E TPGS and Oc-40 at a particular wt% ratio (4.5:2.0) produced excellent drug entrapment, loading, small mixed nanomicellar size and narrow PDI. Solubility of DEX in MNF is improved by~6.3-fold relative to normal aqueous solubility. Critical micellar concentration (CMC) for blend of polymers (4.5:2.0) was found to be lower (0.012 wt%) than the individual polymers (Vit E TPGS (0.025 wt%) and Oc-40 (0.107 wt%)). No significant effect on mixed nanomicellar size and PDI with one-factor or multifactor interactions was observed. Qualitative 1 H NMR studies confirmed absence of free drug in the outer aqueous MNF medium. MNF appeared to be highly stable. Cytotoxicity studies on rabbit primary corneal epithelial cells did not indicate any toxicity suggesting MNF of dexamethasone is safe and suitable for human topical ocular drops after further in vivo evaluations.

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Section: Entrapment and Loading Efficiencymentioning

confidence: 99%

Optimization of Dexamethasone Mixed Nanomicellar Formulation

et al. 2014

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“…19 The detailed studies have indicated that thermodynamically stable reverse micelles were formed with a mixture of Triton X-45 and Span 80 in n-hexane at total concentrations lower than 60 mmol/L and molar fractions of Triton X-45 less than 0.6. Characterization of the system indicated that 30 mmol/L of the surfactants at equimolar Triton X-45 and Span 80 had water content (the molar ratio of water to total surfactant concentration in the organic phase) of about 25 and gave high protein solubilization capacity.…”

Section: Nickel-chelate Reverse Micellesmentioning

confidence: 99%

“…The water content kept nearly unchanged at extensive pH values (3)(4)(5)(6)(7)(8)(9)(10)(11) and chelated metal concentration range. 19 Besides the nonionic surfactants, the organic phase contained 0 to 8 mmol/L sodium neutralized HDEHP (NaDEHP), which was prepared as described previously. 19 The organic phase was contacted by mixing with 50 mL of 50 mmol/L NiSO 4 in a 250-mL flask by reciprocal shaking in a water bath at 25 C for 60 min.…”

Section: Nickel-chelate Reverse Micellesmentioning

confidence: 99%

“…19 Besides the nonionic surfactants, the organic phase contained 0 to 8 mmol/L sodium neutralized HDEHP (NaDEHP), which was prepared as described previously. 19 The organic phase was contacted by mixing with 50 mL of 50 mmol/L NiSO 4 in a 250-mL flask by reciprocal shaking in a water bath at 25 C for 60 min. The Ni(II)-chelated organic phase was then collected by gravity settling and washed three times with 0.02 mol/L Tris-HCl buffer (pH 8.0) to remove any unbound metal ions.…”

Section: Nickel-chelate Reverse Micellesmentioning

confidence: 99%

“…Recently, the authors developed a new nonionic reverse micellar system composed of two nonionic surfactants (Triton X-45 and Span 80) and di(2-ethylhexyl) phosphoric acid (HDEHP). 19 As a cosurfactant incorporated into the nonionic reverse micelles, HDEHP dimers can chelate transition metal ions such as Cu 2þ and Ni 2þ in a equimolar pattern, leading to the fabrication of metal-chelate affinity-based reverse micelles. The system has been well characterized in its water content, micellar size and aggregation number, and demonstrated to work for selective metal-chelate extraction of histidine-rich protein (myoglobin) from myoglobin and lysozyme mixtures.…”

Section: Introductionmentioning

confidence: 99%

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His‐tagged protein purification by metal‐chelate affinity extraction with nickel‐chelate reverse micelles

Dong

Feng

Sun

2010

Biotechnology Progress

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Di(2-ethylhexyl) phosphoric acid (HDEHP) was used as a transition metal ion chelator and introduced to the nonionic reverse micellar system composed of equimolar Triton X-45 and Span 80 at a total concentration of 30 mmol/L. Ni(II) ions were chelated to the HDEHP dimers in the reverse micelles, forming a complex denoted as Ni(II)R(2). The Ni(II)-chelate reverse micelles were characterized for the purification of recombinant hexahistidine-tagged enhanced green fluorescent protein (EGFP) expressed in Escherichia coli. The affinity binding of EGFP to Ni(II)R(2) was proved by investigation of the forward and back extraction behaviors of purified EGFP. Then, EGFP was purified with the affinity reverse micelles. It was found that the impurities in the feedstock impeded EGFP transfer to the reverse micelles, though they were little solubilized in the organic phase. The high specificity of the chelated Ni(2+) ions toward the histidine tag led to the production of electrophoretically pure EGFP, which was similar to that purified by immobilized metal affinity chromatography. A two-stage purification by the metal-chelate affinity extraction gave rise to 87% recovery of EGFP. Fluorescence spectrum analysis suggests the preservation of native protein structure after the separation process, indicating the system was promising for protein purification.

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Theoretical Aspects and Applications of Aqueous Two‐Phase Systems

Singla

Sit

2022

ChemBioEng Reviews

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The aqueous two-phase system (ATPS) is commonly known as a technique that yields high-purity products in a single step. It is particularly advantageous for purifying biomolecules like proteins, nucleic acids, enzymes, viruses etc. Currently, aqueous two-phase extraction (ATPE), i.e., liquid-liquid extraction, involves the transfer of the solute from one aqueous phase to another. In ATPE, for recovery of biomolecules, polymer-polymer and polymersalt type systems are used. The most recent developments with respect to recovery of biomolecules by ATPS are reviewed and discussed, covering the mechanism, which controls the phase formation, the conditions of solute partitioning in ATPS processes, and factors influencing the ATPS including concentration and molecular weight (MW) of polymers, types of salt, pH, and temperature. In addition, also the increasing applications of ATPS for the recovery of high-value bioproducts, the benefits of the ATPS recovery system, and the recent developments of alternative low-cost ATPS are highlighted.

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A metal‐chelate affinity reverse micellar system for protein extraction

Cited by 16 publications

References 45 publications

Optimization of Dexamethasone Mixed Nanomicellar Formulation

Optimization of Dexamethasone Mixed Nanomicellar Formulation

His‐tagged protein purification by metal‐chelate affinity extraction with nickel‐chelate reverse micelles

Theoretical Aspects and Applications of Aqueous Two‐Phase Systems

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