Synthesis of magnetic nanoparticles with an IDA or TED modified surface for purification and immobilization of poly-histidine tagged proteins

Zeng, Kai; Sun, E; Liu, Zewen; Guo, Junhui; Yuan, Chengqing; Yang, Ying; Xie, Hao

doi:10.1039/c9ra10473a

Cited by 11 publications

(11 citation statements)

References 82 publications

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“…where vt is the sedimentation rate, r is the distance of the particle from the axis of rotation, M the molar mass of the particle and ω is the angular velocity of the rotor equation (6). S serves to normalize the vt of a particle by the acceleration applied to it (r. ω 2 ).…”

Section: (7)mentioning

confidence: 99%

“…Within the vast alternatives of biological systems for recombinant protein production, E. coli is the preferred microorganism for protein expression because of the easy handling and storage 4 . Besides, in the protein purification step, histidine tags (His-tag) have gained great popularity 5,6 . His-tags (typically containing six or more consecutive histidine residues) may be used for protein purification by immobilized metal affinity chromatography (IMAC), where divalent cations (usually Ni 2+ or Co 2+ ) are adsorbed in an agarose matrix.…”

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confidence: 99%

“…The viscosity and density values (required for S calculations) in the case of β-Gal solutions were determined using SEDNTERP software 29. S has the dimensions of time units expressed in Sverdberg (1S=10 −13 s) and can be calculated by equations(6) and(7)…”

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confidence: 99%

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Advantages and Disadvantages of His-Tagged Beta-Galactosidase

Flores

Clop

Barra

et al. 2020

Preprint

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β-Galactosidase is one of the most important biotechnological enzyme used in the dairy industry, pharmacology and in molecular biology. In our laboratory we have overexpressed a recombinant β-galactosidase in Escherichia coli (E. coli). This enzyme differs from its native version (β-GalWT) in that 6 histidine residues have been added to the carboxyl terminus in the primary sequence (β-GalHis), which allows its purification by immobilized metal affinity chromatography (IMAC). In this work we compared the functionality and structure of both proteins and evaluated their catalytic behavior on the kinetics of lactose hydrolysis. We observed a significant reduction in the enzymatic activity of β-GalHis with respect to β-GalWT. Although, both enzymes showed a similar catalytic profile as a function of temperature, β-GalHis presented a higher resistance to the thermal inactivation and evidenced greater half-life time compared to β-GalWT. At room temperature, β-GalHis showed a fluorescence spectrum compatible with a partially unstructured protein however, it exhibited a lower tendency to the thermal-induced unfolding with respect to β-GalWT. Analytical ultracentrifugation experiments demonstrated that the population of β-GalHis molecules exhibited a higher proportion of monomers and a lower proportion of tetrameric species with respect to the His-tag free protein. The impairment of tetramerization may would explain the negative effect of the presence of His-tag on the enzymatic activity. In addition, the present results, analyzed in the context of the available literature, suggest that the effect of the His-tag is protein-specific.

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Section: (7)mentioning

confidence: 99%

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confidence: 99%

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Advantages and Disadvantages of His-Tagged Beta-Galactosidase

Flores

Clop

Barra

et al. 2020

Preprint

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“…At present, several materials have been introduced to purify His‐tagged protein. Metal‐resin complex is mostly obtained support by fixing metal ions on resin microspheres through complex agents, such as nitrogen triacetic acid (NTA) [10, 11], or iminodiacetic acid (IDA) [12, 13]. However, these materials have some drawbacks like time‐consuming operation, complicated pretreatment, poor mechanical stability [14, 15].…”

Section: Introductionmentioning

confidence: 99%

“…Furthermore, protein tags also play an essential role in the purification of the recombinant protein. Due to the relatively small [12,13]. However, these materials have some drawbacks like time-consuming operation, complicated pretreatment, poor mechanical stability [14,15].…”

Section: Introductionmentioning

confidence: 99%

One‐step selective affinity purification and immobilization of His‐tagged enzyme by recyclable magnetic nanoparticles

Zhou

et al. 2021

Engineering in Life Sciences

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The NiFe2O4 magnetic nanoparticles (NF‐MNPs) were prepared for one‐step selective affinity purification and immobilization of His‐tagged recombinant glucose dehydrogenase (GluDH). The prepared nanoparticles were characterized by a Fourier‐transform infrared spectrophotometer and microscopy. The immobilization and purification of His‐tagged GluDH on NF‐MNPs were investigated. The optimal immobilization conditions were obtained that mixed cell lysis and carriers in a ratio of 0.13 in pH 8.0 Tris‐HCl buffer at 30℃ and incubated for 2 h. The highest activity recovery and protein bindings were 71.39% and 38.50 μg mg–1 support, respectively. The immobilized GluDH exhibited high thermostability, pH‐stability and it can retain more than 65% of the initial enzyme after 10 cycles for the conversion of glucose to gluconolactone. Comparing with a commercial Ni‐NTA resin, the NF‐MNPs displayed a higher specific affinity with His‐tagged recombinant GluDH.

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Aspects of high-performance and bio-acceptable magnetic nanoparticles for biomedical application

Kush

Kumar

Singh

et al. 2021

Asian Journal of Pharmaceutical Sciences

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This review covers extensively the synthesis & surface modification, characterization, and application of magnetic nanoparticles. For biomedical applications, consideration should be given to factors such as design strategies, the synthesis process, coating, and surface passivation. The synthesis method regulates post-synthetic change and specific applications in vitro and in vivo imaging/diagnosis and pharmacotherapy/administration. Special insights have been provided on biodistribution, pharmacokinetics, and toxicity in a living system, which is imperative for their wider application in biology. These nanoparticles can be decorated with multiple contrast agents and thus can also be used as a probe for multi-mode imaging or double/triple imaging, for example, MRI-CT, MRI-PET. Similarly loading with different drug molecules/dye/fluorescent molecules and integration with other carriers have found application not only in locating these particles in vivo but simultaneously target drug delivery/hyperthermia inside the body. Studies are underway to collect the potential of these magnetically driven nanoparticles in various scientific fields such as particle interaction, heat conduction, imaging, and magnetism. Surely, this comprehensive data will help in the further development of advanced techniques for theranostics based on high-performance magnetic nanoparticles and will lead this research area in a new sustainable direction.

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Synthesis of magnetic nanoparticles with an IDA or TED modified surface for purification and immobilization of poly-histidine tagged proteins

Abstract: This article reports a novel approach for synthesizing magnetic nanoparticles with a modified surface for purification and immobilization of histidine-tagged proteins.

Cited by 11 publications

References 82 publications

Advantages and Disadvantages of His-Tagged Beta-Galactosidase

Advantages and Disadvantages of His-Tagged Beta-Galactosidase

One‐step selective affinity purification and immobilization of His‐tagged enzyme by recyclable magnetic nanoparticles

Aspects of high-performance and bio-acceptable magnetic nanoparticles for biomedical application

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