Synthesis of magnetic nanoparticles functionalized with histidine and nickel to immobilize His-tagged enzymes using β-galactosidase as a model

Andrade, Bruna Coelho de; Gennari, Adriano; Renard, Gaby; Nervis, Brenda Da Rolt; Benvenutti, Edílson V.; Costa, Tânia Maria Haas; Nicolodi, Sabrina; Silveira, Nádya Pesce da; Chies, Jocelei Maria; Volpato, Giandra; Souza, Cláucia Fernanda Volken de

doi:10.1016/j.ijbiomac.2021.06.060

Cited by 22 publications

(8 citation statements)

References 73 publications

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“…The determined magnetization of BION and the only slight decrease by around 5 emu g –1 upon protein adsorption are consistent with already published data. , Also, the strong shielding of the magnetization due to the coating of ION@TEOS agrees with the literature. , The larger intrinsic magnetization of BION and BION@rSpA likely contributed to the generally faster determined magnetophoretic sedimentation velocities compared to ION@TEOS (Figure E). In addition, the magnetophoretic velocity strongly correlates with the hydrodynamic diameter. , Thus, the increased agglomeration seen for the uncoated particles probably also benefited the fast magnetophoretic attraction.…”

Section: Resultssupporting

confidence: 91%

Direct Affinity Ligand Immobilization onto Bare Iron Oxide Nanoparticles Enables Efficient Magnetic Separation of Antibodies

Zimmermann,

Kaveh-Baghbaderani,

Eilts

et al. 2024

ACS Appl. Bio Mater.

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Magnetic separation is a promising alternative to chromatography for enhancing the downstream processing (DSP) of monoclonal antibodies (mAbs). However, there is a lack of efficient magnetic particles for successful application. Aiming to fill this gap, we demonstrate the suitability of bare iron oxide nanoparticles (BION) with physical site-directed immobilization of an engineered Protein A affinity ligand (rSpA) as an innovative magnetic material. The rSpA ligand contains a short peptide tag that enables the direct and stable immobilization onto the uncoated BION surface without commonly required laborious particle activation. The resulting BION@rSpA have beneficial characteristics outperforming conventional Protein A-functionalized magnetic particles: a simple, fast, low-cost synthesis, a particle size in the nanometer range with a large effective specific surface area enabling large immunoglobulin G (IgG) binding capacity, and a high magnetophoretic velocity advantageous for fast processing. We further show rapid interactions of IgG with the easily accessible rSpA ligands. The binding of IgG to BION@rSpA is thereby highly selective and not impeded by impurity molecules in perfusion cell culture supernatant. Regarding the subsequent acidic IgG elution from BION@rSpA@IgG, we observed a hampering pH increase caused by the protonation of large iron oxide surfaces after concentrating the particles in 100 mM sodium acetate buffer. However, the pH can be stabilized by adding 50 mM glycine to the elution buffer, resulting in recoveries above 85% even at high particle concentrations. Our work shows that BION@rSpA enable efficient magnetic mAb separation and could help to overcome emerging bottlenecks in DSP.

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Section: Resultssupporting

confidence: 91%

Direct Affinity Ligand Immobilization onto Bare Iron Oxide Nanoparticles Enables Efficient Magnetic Separation of Antibodies

Zimmermann,

Kaveh-Baghbaderani,

Eilts

et al. 2024

ACS Appl. Bio Mater.

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“…Biocatalysts are susceptible to various environmental pressures (high temperature, pH, organic reagents, and ionic strength). Hence, immobilized enzymes are often expected to have excellent temperature and pH stability. − As shown in Figure E,F, the thermal stabilities of both single CLEAs were significantly enhanced compared to free enzymes. Especially for Rha1, after pre-treatment at 80 °C for 2 h, the enzyme activity of free Rha1 was completely lost, whereas the relative activity of Rha1-CLEAs was still 54.19% of its maximum activity.…”

Section: Resultsmentioning

confidence: 97%

Co-Immobilizing Two Glycosidases Based on Cross-Linked Enzyme Aggregates to Enhance Enzymatic Properties for Achieving High Titer Icaritin Biosynthesis

Liu

Wei

Cheng

et al. 2022

J. Agric. Food Chem.

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Icaritin is a rare and high-value isopentane flavonoid compound with remarkable activities. Increasing yields while reducing cost has been a great challenge in icaritin production. Herein, we first reported a high titer icaritin biosynthesis strategy from epimedin C through co-immobilizing α-l-rhamnosidase (Rha1) and β-glucosidase (Glu4) using cross-linked enzyme aggregates (CLEAs). The created CLEAs exhibited excellent performances in terms of catalytic activity, thermal stability, pH stability, and reusability. Notably, Rha1-CLEAs (K i : 1 M) and Glu4-CLEAs (K i : 0.1 M) were more tolerant to sugars (glucose or rhamnose) than free enzymes (0.1 M for Rha1 and 0.007 M for Glu4) by immobilization, achieving the highest icaritin productivity under the highest substrate concentration ever reported. Finally, about 34.24 g/L icaritin could be obtained from 100 g/L epimedin C within 8 h, indicating the great potential for industrialization. This study also provides a promising strategy for the low-cost production of other high-value aglycone compounds by solving poor stability and sugar inhibition of glycosidase.

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“…Based on the Arrhenius method, the deactivation energy (Eq. (4) ) was obtained with the following equation [ 45 ]:

where k d is the deactivation rate constant, A is a constant, E d is the deactivation energy, R (8.314 × 10 −3 kJ mol −1 K −1 ) is the universal gas constant, and T is the absolute temperature. The time required for the enzyme to show a decrease to 50% of the initial activity (half-life) is t 1/2 = ln 2/ k .…”

Section: Resultsmentioning

confidence: 99%

“…(6) ) increased slightly from 102.54 to 103.22 kJ/mol for the immobilized enzyme. This demonstrated that immobilization improved the thermal stability of the enzyme, making it more resistant to denaturation and conformational changes [ 47 ]. As seen in Eq.…”

Section: Resultsmentioning

confidence: 99%

Nanocatalytic performance of pectinase immobilized over in situ prepared magnetic nanoparticles

Navarro-López,

Bautista-Ayala,

Rosales-De la Cruz

et al. 2023

Heliyon

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Synthesis of magnetic nanoparticles functionalized with histidine and nickel to immobilize His-tagged enzymes using β-galactosidase as a model

Cited by 22 publications

References 73 publications

Direct Affinity Ligand Immobilization onto Bare Iron Oxide Nanoparticles Enables Efficient Magnetic Separation of Antibodies

Direct Affinity Ligand Immobilization onto Bare Iron Oxide Nanoparticles Enables Efficient Magnetic Separation of Antibodies

Co-Immobilizing Two Glycosidases Based on Cross-Linked Enzyme Aggregates to Enhance Enzymatic Properties for Achieving High Titer Icaritin Biosynthesis

Nanocatalytic performance of pectinase immobilized over in situ prepared magnetic nanoparticles

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