APPLIED PHYSICS A-MATERIALS SCIENCE & PROCESSING
·
2019
·Vol. 125
·期 5
·1-10
·
IF 1.784
Guo Huiling,Wang Wenjing,Zhou Fengzhen
In this study, core–shell Fe 3 O 4 @Au nanoparticles with good chemical stability and tunable particle size were synthesized via Au–S bonding, which overcomes the challenges encountered for the reported preparation of Fe 3 O 4 @Au. The surface of nanoparticles was investigated by TEM, XRD, XPS, FT-IR, VSM and DLS which characterized the size, morphology, chemical lattice, elemental analysis, functional groups, magnetic strength, and size distribution. The results showed that Fe 3 O 4 @Au/NTA-Ni 2+ magnetic nanocomposites were spherical with an average diameter of 469.5 nm, and superparamagnetic with saturation magnetization of 7 emu/g. The magnetic nanocomposites were directly utilized for one-step purification of His-tagged proteins from Description Escherichia coli lysate. The as-separated proteins were qualitatively validated by gel analysis and quantitatively measured by UV–visible spectroscopy. The binding capacity of Fe 3 O 4 @Au/NTA-Ni 2+ towards His-tagged proteins was 48.3 mg/g. The target proteins were separated with high purity and separation efficiency up to 96.6%. Their specificity and enrichment ability were maintained during six cycles of adsorption–elution operations. The advantages of this novel nanoparticle can open a window for the multifunction of nanomaterials and their biological application.
研究领域:
生物化学
·
纳米技术
·
蛋白质工程
✓ 已验证
产品关联
DOI
高影响因子
JOURNAL OF SEPARATION SCIENCE
·
2018
·Vol. 42
·期 3
·744-753
·
IF 2.415
Mengmeng Chang,Qian Qin,Bohong Wang,Tian Xia,Wangjie lv,Xiaoshan Sun,Xianzhe Shi,Guowang Xu
Employing immobilized metal-ion affinity chromatography and magnetic separation could ideally provide a useful analytical strategy for purifying His-tagged protein. In the current study, a facile route was designed to prepare CMPEI-Ni 2+ @SiO 2 @Fe 3 O 4 (CMPEI=carboxymethylated
polyethyleneimine) magnetic nanoparticles composed of a strong magnetic core of Fe 3 O 4 and a Ni 2+ -immobilized carboxymethylated polyethyleneimine coated outside shell, which was formed by electrostatic interactions between polyanionic electrolyte of carboxymethylated polyethyleneimine and positively charged surface of 3-(trimethoxysilyl)propylamin modified SiO 2 @Fe 3 O 4 . The resulting CMPEI-Ni 2+ @SiO 2 @Fe 3 O 4 composite nanoparticles displayed well-uniform structure and high magnetic responsiveness. Hexa His-tagged peptides and purified His-tagged recombinant retinoid X receptor alpha were chosen as the model samples to evaluate the adsorption, capacity, and reusability of the composite nanoparticles. The results demonstrated the CMPEI-Ni 2+ @SiO 2 @Fe 3 O 4 nanoparticles possessed rapid adsorption, large capacity, and good recyclability. The obtained nanoparticles were further used to purify His-tagged protein in practical environment. It was found that the nanoparticles could selectively capture His-tagged recombinant retinoid X receptor protein from complex cell lysate. Owing to its easy synthesis, large binding capacity, and good reusability, the prepared CMPEI-Ni 2+ @SiO 2 @Fe 3 O 4 magnetic nanoparticles have great potential for application in biotechnological fields.
研究领域:
生物化学
·
纳米技术
·
蛋白质工程
✓ 已验证
DOI
PubMed
高影响因子
Nanoscience and Nanotechnology Letters
·
2018
·Vol. 10
·期 3
·435-439
·
IF 2.917
Xu Hongpan,Sun Tingting,Huang Yuanfu,Song Lian,Li Zhiyang,Wang Wei
Placing of affinity tags at desired position of the proteins has been widely used to facilitate protein separation. One of the most commonly method for protein separation is nickel ion (Ni2+) affinity chromatography. Complex steps and time consuming in nickel ion affinity
chromatography have caused high demand for alternative purification methods that allow direct and rapid separation of His-tagged proteins. In the present study, magnetic core–shell beads enriched with Ni2+-nitrilotriacetate (Ni-NTA) on their surface were prepared in this study
by precipitation polymerization, with a uniform size of about 10 μm. The presented Ni2+ amount was 2.23%, which endowed the spheres with excellent magnetic responsivity and dispersity. We compared the magnetic beads enrichment method and nickel ion affinity chromatography
method for extraction of His-tagged protein sample with different concentrations. The results revealed that, when compared with nickel ion affinity chromatography, extraction efficiency, purity of the extracted protein, and regeneration effort were much higher with the magnetic beads enrichment
method. By using magnetic beads enrichment method, samples could be separated within 20 s, and purified within 1 h, with unlimited flow velocity and sample capacity. In addition, the magnetic beads enrichment method was particularly promising for the separation of His-tagged proteins in less
samples.
研究领域:
生物化学
·
蛋白质组学
✓ 已验证
DOI
高影响因子
BIOCHEMICAL ENGINEERING JOURNAL
·
2020
·
IF 3.475
Yongxing Li,Pengqian Luan,Liya Zhou,Saiguang Xue,Yihang Liu,Yunting Liu,Yanjun Jiang,Jing Gao
The synthesis or decomposition of complex substances through the combination of metals and enzymes is desirable but still a huge challenge. A kind of yolk-shell structured Co/C@SiO 2 @Ni/C nanocomposites were prepared by the polyhedral ZIF-67@SiO 2 composite as a precursor through the extended stöber method and subsequent carbonization treatment. The SiO 2 as an intermediate protective layer stabilized the morphology and structure of the carbonized products. The metallic nickel nanoparticles were uniformly distributed on the surface of Co/C@SiO 2 @Ni/C for highly specific absorption of His-tagged organophosphohydrolase (OpdA) from cell lysates via affinity, and therefore an integrated nanocatalyst (INC) named OpdA@Co/C@SiO 2 @Ni/C was successfully prepared. The prepared INC was applied to achieve a one-pot chemoenzymatic cascade degradation of organophosphates (OPs) to p -aminophenol (PAP) within 30 min. In addition, the INC proved to be a simple and high-sensitivity nanocatalyst for the detection of methyl-parathion with a low limit of detection (0.30 μM). The INC exhibited improved pH stability, thermal stability, SDS resistance, and storage stability compared to that of free enzymes. When OpdA@Co/C@SiO 2 @Ni/C was reused for 7 cycles, the cascade degradation efficiency of methyl-parathion could retain more than 60 %. This study provides a strategy for constructing a multifunctional nanocatalyst, and further demonstrated its potential in biocatalysis, accurate detection of OPs and remediation of environmental pollution.
研究领域:
生物化学
·
环境科学
·
材料科学
✓ 已验证
产品关联
DOI
高影响因子
BIOSENSORS & BIOELECTRONICS
·
2013
·
IF 5.437
Huanshun Yin,Bing Sun,Yunlei Zhou,Mo Wang,Zhenning Xu,Zhengliang Fu,Shiyun Ai
In this work, we fabricated a novel photoelectrochemical immunosensor for assay of DNA methylation , where Bi 2 S 3 nanorods were used as photoelectric conversion material, MBD1 protein (a kind of methyl bonding domain protein) was used as DNA methylation recognizing unit, anti-his tag antibody was used to further inhibit the photocurrent and increase the detection sensitivity. The results demonstrated that Bi 2 S 3 possessed excellent photoelectron property. The detection conditions, such as Bi 2 S 3 concentration, MBD1 protein concentration, incubation time of MBD1 protein, antibody concentration and antibody incubation time, were optimized. Under optimal experimental conditions, the photocurrent variation was proportional to the logarithm of methylated target DNA concentration from 10 −9 to 10 –13 M with detection limit of 3.5×10 –14 M ( S / N =3). Moreover, the immunosensor presented high detection specificity, even distinguishing single-base mismatched sequence.
研究领域:
生物化学
·
分子生物学
✓ 已验证
产品关联
DOI
PubMed
高影响因子
APPLIED MICROBIOLOGY AND BIOTECHNOLOGY
·
2014
·Vol. 98
·期 12
·5665-5675
·
IF 3.811
Xu Weiyuan,Cao Huazhen,Ren Guangwei,Xie Hujun,Huang Jianying,Li Shijun
A sorbent based on affinity ionic liquid (AIL), triazacyclononane-ionic liquid, was synthesized, characterized, and applied to the extraction of histidine (His)-tagged proteins from aqueous buffer to ionic liquid (IL) phase. The adsorbed His-tagged proteins could be back-extracted from the IL phase to the aqueous buffer with an imidazole solution. The specific binding of His-tagged proteins with AIL/IL could be affected by a few factors including the ionic strength and coordinated metal ions. In the case of His-tagged enhanced green fluorescent protein (EGFP), the maximum binding capacity of Cu 2+ -AIL/IL reached 2.58 μg/μmol under the optimized adsorption conditions. The eluted His-tagged EGFP kept fluorescent and remained active through the purification process. Moreover, a tandem extraction process successively using Cu 2+ -AIL/IL and Zn 2+ -AIL/IL systems was developed, which was proven very efficient to obtain the ultimate protein with a purity of about 90 %. An effective reclamation method for the AIL/IL extraction system was further established. The sorbent could be easily regenerated by removing metal ions with EDTA and the followed reimmobilization of metal ions. Easy handling of the presented M 2+ -AIL/IL system and highly specific ability to absorb His-tagged proteins make it attractive and potentially applicable in biomolecular separation.
研究领域:
生物化学
·
蛋白质工程
✓ 已验证
产品关联
DOI
PubMed
高影响因子
RSC Advances
·
2020
·Vol. 10
·期 39
·23270-23275
·
IF 3.119
Xueyan Zou,Yu Zhang,Jinqiu Yuan,Zhibo Wang,Rui Zeng,Kun Li,Yanbao Zhao,Zhijun Zhang
In this study, porous silica nanoparticles functionalized with a thiol group (SiO2–SH NPs) were synthesized via a one-pot method. Subsequently, iminodiacetic acid was modified, and further adsorption of Ni2+ ions was conducted to obtain a SiO2–S/NH–Ni nano-adsorbent. Then, transmission electron microscopy (TEM), scanning electron microscopy (SEM), Fourier transform infrared (FT-IR) spectroscopy, thermogravimetric analysis (TG) and X-ray diffraction (XRD) were employed to characterize its morphology and composition. The results indicate that the SiO2–S/NH–Ni nano-adsorbent is porous, has an average diameter of 77.1 nm and has a small porous structure of about 3.7 nm in the silica skeleton. The Brunauer–Emmett–Teller (BET) surface area and total pore volume were 537.2 m2 g−1 and 3.3 cm3 g−1, respectively, indicating a large BET surface area. The results indicate that the as-prepared SiO2–S/NH–Ni nano-adsorbent would be suitable to selectively and efficiently bind His-tagged proteins from an E. coli cell lysate. The SDS-PAGE results show that the as-prepared nano-adsorbent presents specifically to both His-tagged CPK4 and His-tagged TRX proteins, indicating the nano-adsorbent can be used to effectively separate His-tagged proteins and is universal to all His-tagged fusion proteins. We also found that the as-prepared nano-adsorbent exhibits a low detection limit (1.0 × 10−7 mol L−1) and a strong regeneration ability based on four regeneration experiments that were particularly suited to the separation of His-tagged proteins.
研究领域:
生物化学
·
纳米材料科学
·
蛋白质组学
✓ 已验证
DOI
高影响因子
MICROCHIMICA ACTA
·
2015
·Vol. 183
·期 2
·683-688
·
IF 3.741
Jia Shengsong,Li Pengfei,Koh Kwangnak,Chen Hongxia
The article describes the use of a designed histidine (His)-tagged arginine-glycine-aspartic acid (RGD) peptide as a linker to bind integrin to captured cancer cells, and how to amplify the surface plasmon resonance (SPR) signal after binding of NiO nanoparticles (NiO-NPs) via the His-tag on the peptide. Specifically, breast cancer cells were captured via the interaction between human mucin (MUC-1) and a gold surface modified with a MUC-selective aptamer. The resulting cytosensor exhibits specificity and high sensitivity which is due to the enhancement of the SPR signal by NiO-NPs. The breast cancer cell line MCF-7 can be easily distinguished from normal islet beta cells by using this biosensor. Implementation of the His-tagged RGD peptide modified with NiO-NPs resulted in 20-fold enhancement of the SPR signal at the limit of detection. Hence, the actual limit of detection is lowered to 136 cells per mL. In our perception, this cytosensor has a large clinical potential in that it may also be used to detect various other kinds of tumor cells.
Histidine-tagged arginine-glycine-aspartic acid peptide carried by NiO interacts with the integrin of cancer cells and amplifies the SPR signal resulting from interaction between human mucin (MUC-1) and a gold surface modified with a MUC-selective aptamer. As little as 136 cells·mL −1 are detectable.
研究领域:
生物传感技术
·
肿瘤检测
·
医学诊断
✓ 已验证
产品关联
DOI
高影响因子
JOURNAL OF WUHAN UNIVERSITY OF TECHNOLOGY-MATERIALS SCIENCE EDITION
·
2018
·Vol. 33
·期 3
·559-565
·
IF 0.524
Guo Huiling,Sun Honghao,Su Zhengding,Hu Shuchao,Wang Xiaotao
A facile approach has been developed to synthesize Fe 3 O 4 @PAM (polyacrylamide) nanoparticles (NPs) with carboxyl groups on the surfaces by copolymerization with acrylamide and acrylic acid in Fe 3 O 4 NPs aqueous suspension. Nitrilotriacetic acid (NTA) was conjugated to the magnetic NPs via well-known carboniimide chemistry using EDC and NHS. The Ni 2+ ions loaded on the surface of NPs provide abundant docking sites for immobilization of His-tagged green fluorescent proteins (His-tagged GFP). The high magnetic property of Fe 3 O 4 @PAM@NTA-Ni 2+ allows an easy separation of the NPs from solution under an external magnetic field, with high His-tagged protein binding capacity (42 μg protein/mg of NPs). The NPs can be recycled for at least four times without significant loss of binding capacity to proteins. These materials show great potential to separate His-tagged protein with low-cost purification at industrial scale.
研究领域:
材料科学
·
生物化学
·
生物技术
·
纳米技术
✓ 已验证
产品关联
DOI
高影响因子
IET Nanobiotechnology
·
2019
·Vol. 14
·期 1
·67-72
·
IF 1.925
Wenjing Wang,Fengzhen Zhou,Xiyao Cheng,Zhengding Su,Huiling Guo
To effective capture and universal enrichment of His-tagged protein, polyacrylic acid (PAA) brushes were used to encapsulate Fe 3 O 4 nanoparticles, connect NTA, and Ni 2+ to prepare magnetic beads. These materials provide many advantages, such as excellent stability, tuneable particle size, and a surface for further functionalisation with biomolecules. His-tagged green fluorescence protein (GFP) was separated efficiently, and the binding capacity of Fe 3 O 4 /MPS@PAA/NTA-Ni 2+ was 93.4 mg/g. Compared with High-Affinity Ni-NTA Resin and Ni-NTA Magnetic Agarose Beads, Fe 3 O 4 /MPS@PAA/NTA-Ni 2+ nanocomposites exhibited higher separation efficiency and binding capacity towards His-tagged GFP. Moreover, the selectivity and recyclability of them for the target proteins were maintained well after six cycles. This study would widen the application of PAA in constructing multifunctional nanocomposites for biomedical fields.
研究领域:
生物技术
·
蛋白质工程
·
纳米材料
✓ 已验证
产品关联
DOI
PubMed
高影响因子
ENZYME AND MICROBIAL TECHNOLOGY
·
2022
·
IF 3.705
Liya Zhou,Yaping Ouyang,Weixi Kong,Teng Ma,Hao Zhao,Yanjun Jiang,Jing Gao,Li Ma
In this study, a novel kind of Ni-NTA modified monodispersed SiO 2 nanoflowers (Ni-NTA@SiO 2 nanoflowers) were successfully synthesized. The obtained Ni-NTA@SiO 2 nanoflowers were used to specifically adsorb and purify His-tagged old yellow enzyme (OYE1) and glucose dehydrogenase (GDH), which allows access to optically pure (3 S)− 3-methyl-cyclohexanone through asymmetric hydrogenation reaction, and forms a cofactor regeneration system. The protein loading amount on Ni-NTA@SiO 2 nanoflowers was 40.17 mg/g support and the activity recoveries of OYE1 and GDH were 81.53% and 79.68%, respectively. The effects of pH and temperature on the activity of free and co-immobilized enzymes were investigated, and the stability as well as reusability were also measured. Compared to free enzymes, the co-immobilized enzymes showed higher thermal and storage stability. The co-immobilized enzymes were applied to asymmetric reduction of C C bonds for the synthesis of a chiral center with excellent enantioselectivity (ee > 99%), and the conversion was 46.02% after 7 cycles. This work introduced a one-pot multi-enzyme purification and co-immobilization strategy to construct efficient cofactor regeneration system with high activity and stability.
研究领域:
生物化学
·
酶工程
·
纳米材料科学
✓ 已验证
产品关联
DOI
PubMed
高影响因子
ACS Applied Materials & Interfaces
·
2013
·
IF 5.008
Yuting Zhang,Yongkun Yang,Wanfu Ma,Jia Guo,Yao Lin,Changchun Wang
A facile approach has been developed to synthesize Fe3O4/PMG (poly (N,N′-methylenebisacrylamide-co-glycidyl methacrylate)) core/shell microspheres using distillation–precipitation polymerization. Treating PMG shell with iminodiacetic acid (IDA) and Ni2+ yields composite microspheres of Fe3O4/PMG/IDA–Ni2+. The Ni2+ ions loaded on the surface of microspheres provide abundant docking sites for immobilization of histidine-tagged proteins. The high saturation magnetization of Fe3O4/PMG (23 emu/g), determined by vibrating sample magnetometer (VSM), allows an easy separation of the microspheres from solution under an external magnetic field. The composite microspheres were used to purify two His-tagged cellulolytic enzymes (Cel48F and Cel9G) directly from crude cell lysates with high binding affinity, capacity, and specificity. The microspheres can be recycled for many times without significant loss of binding capacity to enzymes. The immobilized enzymes on the surface of microspheres well retain their biological activities in degradation of cellulose. These materials show great potential in the biomedical and biotechnological applications that require low-cost purification of recombinant proteins and instant enzyme immobilization at an industrial scale.
研究领域:
生物技术
·
生物化学
·
材料科学
·
酶学
✓ 已验证
产品关联
DOI
PubMed
高影响因子
危险品化学品经营许可证（带存储）