Microgels for Efficient Protein Purification

Mizrahi, Boaz; Irusta, Silvia; McKenna, Marshall; Ştefănescu, Cristina; Yedidsion, Liron; Myint, MyatNoeZin; Langer, Robert; Kohane, Daniel S.

doi:10.1002/adma.201101258

Cited by 20 publications

(20 citation statements)

References 28 publications

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“…Interestingly, after the elution process, the IENFAs could return to their original state without structural failure, indicating the excellent structural stability and reusability of IENFAs. Moreover, as compared with the complicated electrophoretic bands of egg white solution, the obtained effluent showed a single and clear electrophoretic band around 14.7 kD of lysozyme (inset of Figure e) . Meanwhile, the extracted protein could be precipitated out by adjusting the buffer pH of to 10.7 (isoelectric point of lysozyme; Figure S40, Supporting Information), demonstrating the effective extraction and purification of IENFAs toward the positively charged lysozyme from egg white containing various negatively charged proteins.…”

Section: Resultsmentioning

confidence: 99%

Highly Carboxylated, Cellular Structured, and Underwater Superelastic Nanofibrous Aerogels for Efficient Protein Separation

Duan

et al. 2019

Adv Funct Materials

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Chromatographic media with synchronously large protein adsorption capacity and high processing flux are highly desired in protein separation; however, the creation of such materials still faces enormous challenges. Herein, a robust strategy to develop highly carboxylated monolithic media by combining nanofibrous aerogels' forming technique and an in situ modification approach is reported. The obtained ion-exchange nanofibrous aerogels (IENFAs) exhibit a unique cellular structure consisting of flexible ceramic nanofibers and a functional polymer wrapping layer, endowing them with outstanding underwater superelasticity and compressive fatigue resistance (nearly no plastic deformation after 1000 compressive cycles). Benefiting from the interconnected nanofibrous cellular structure, good hydrophilicity, high carboxylation, and excellent mechanical properties, the IENFAs exhibit synchronously promoted static (2.9 × 10 3 mg g −1 ) and dynamic (1.7 × 10 3 mg g −1 ) lysozyme adsorption capacities and improved buffer flux (2.17 × 10 4 L m −2 h −1 , gravity driven), which are superior to these reported nanofibrous materials and commercial ion-exchange membranes. The IENFAs also possess outstanding performance stability, easy operation, and excellent regenerability. Moreover, the IENFA-packed column could directly and continuously separate lysozyme from egg white solely driven by gravity, highlighting their excellent practical application performance. This work may provide a new avenue to design and develop next-generation high-performance chromatographic media for bioseparation.

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Section: Resultsmentioning

confidence: 99%

Highly Carboxylated, Cellular Structured, and Underwater Superelastic Nanofibrous Aerogels for Efficient Protein Separation

Duan

et al. 2019

Adv Funct Materials

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“…We identify two major prerequisites for efficient protein immobilization. First, a porous microgel is favored over simple commercial, pre‐functionalized agarose‐based particles as an experimental platform . That way, protein accumulation not only occurs on the particle surface but also throughout the microgel matrix, thus enabling to explore the benefits of a tailored 3D‐microenvironment with defined porosity for optimal protein activity and folding characteristics.…”

Section: Resultsmentioning

confidence: 99%

Cell‐Free Protein Synthesis in Bifunctional Hyaluronan Microgels: A Strategy for In Situ Immobilization and Purification of His‐Tagged Proteins

et al. 2019

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“…In the range of 0.85 < P/P0 < 1.0, a very steep capillary condensation step was observed, indicative of a uniform and large pore size. The total pore volume and pore size of the Ni@MSN support and MLG-Ni@MSN were respectively calculated to be ~0.30 and ~0.36 cm 3 /g, and 16.71 and 21.40 nm (Table S1), which were far larger than that of the reported microgel beads and could enhance the protein yields per unit mass of the carrier [41]. Notably, MLG-Ni@MSN held a larger pore size than Ni@MSN, probably because the To further verify the existence of MLG in artificial, bi-functional nanobiocatalyst, TGA was performed.…”

Section: Characterization Of the Ni@msn Supportmentioning

confidence: 87%

Engineering of a Novel, Magnetic, Bi-Functional, Enzymatic Nanobiocatalyst for the Highly Efficient Synthesis of Enantiopure (R)-3-quinuclidinol

Jiang

et al. 2021

Catalysts

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Ni2+-NTA-boosted magnetic porous silica nanoparticles (Ni@MSN) to serve as ideal support for bi-functional enzyme were fabricated for the first time. The versatility of this support was validated by one-step purification and immobilization of bi-functional enzyme MLG consisting of 3-Quinuclidinone reductase and glucose dehydrogenase, which can simultaneously catalyze both carbonyl reduction and cofactor regeneration, to fabricate an artificial bi-functional nanobiocatalyst (namely, MLG-Ni@MSN). The enzyme loading of 71.7 mg/g support and 92.7% immobilization efficiency were obtained. Moreover, the immobilized MLG showed wider pH and temperature tolerance and greater storage stability than free MLG under the same conditions. The nanosystem was employed as biocatalyst to accomplish the 3-quinuclidinone (70 g/L) to (R)-3-quinuclidinol biotransformation in 100% conversion yield with >99% selectivity within 6 h and simultaneous cofactor regeneration. Furthermore, the immobilized MLG retained up to 80.3% (carbonyl reduction) and 78.0% (cofactor regeneration) of the initial activity after being recycled eight times. In addition, the MLG-Ni@MSN system exhibited almost no enzyme leaching during biotransformation and recycling. Therefore, we have reason to believe that the Ni@MSN support gave great promise for constructing a new biocatalytic nanosystem with multifunctional enzymes to achieve some other complex bioconversions.

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Microgels for Efficient Protein Purification

Cited by 20 publications

References 28 publications

Highly Carboxylated, Cellular Structured, and Underwater Superelastic Nanofibrous Aerogels for Efficient Protein Separation

Highly Carboxylated, Cellular Structured, and Underwater Superelastic Nanofibrous Aerogels for Efficient Protein Separation

Cell‐Free Protein Synthesis in Bifunctional Hyaluronan Microgels: A Strategy for In Situ Immobilization and Purification of His‐Tagged Proteins

Engineering of a Novel, Magnetic, Bi-Functional, Enzymatic Nanobiocatalyst for the Highly Efficient Synthesis of Enantiopure (R)-3-quinuclidinol

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