Journal of Materials Chemistry B
·
2016
·Vol. 4
·期 11
·1960-1967
·
IF 4.872
Senwu Li,Kaiguang Yang,Baofeng Zhao,Xiao Li,Lukuan Liu,Yuanbo Chen,Lihua Zhang,Yukui Zhang
Recombinant protein technology occupies an important position in fields including biopharmaceutics, proteomics, structural and functional biology. However, the purification of His-tagged protein, the majority portion of recombinant protein, is seriously hindered by impurities. These impurities, including host proteins with inherent cysteine and histidine-rich regions or metal centers, are usually beyond the purification ability of commonly used IMAC materials. To remove this barrier, a novel purification material was developed through enhancing the selectivity of IMAC by means of surface epitope imprinting using His-tag, the common terminal of His-tagged protein, as the template. Characterizations including TEM, thermogravimetric analysis, X-ray photoelectron spectroscopy, measurement of DLS size and zeta potential were carried out to prove the fabrication of the imprinted shell. Results exhibited a high imprinting factor of 7.1. Besides, the adsorption kinetics were not affected by the surface imprinted shell and could reach adsorption equilibrium within 15 min. Compared with the substrate IMAC, the novel epitope imprinting enhanced IMAC (EI-IMAC) showed an obvious improvement (5% increase of purity) in the selectivity of His-tagged recombinant protein from crude cell lysis.
研究领域:
生物制药
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蛋白质组学
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结构生物学
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功能生物学
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高影响因子
JOURNAL OF CHROMATOGRAPHY B-ANALYTICAL TECHNOLOGIES IN THE BIOMEDICAL AND LIFE SCIENCES
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2021
·
IF 3.205
XinHui Wang,JiaLin Yu,HaoDing Yang,Jian Shen,HaiLong Liu,JiaHong Zhou
Ti-based immobilized metal affinity chromatography (IMAC) nanomaterial has shown high potential in phosphoproteome mass-spectrometric (MS) analysis. However, the limited surface area and poor solubility will greatly restrict its use in phosphoproteome research. To overcome these two key drawbacks, a novel Ti-based IMAC nanomaterial was prepared by Ti-bonded β-cyclodextrin (β-CD) anchored on the surface of carbon nanotubes (CNTs) (denoted as COOH-CNTs-CD-Ti) and successfully applied as a biofunctional adsorbent for selectively enriching trace phosphopeptides. In the selective enrichment process, CNTs provided greater surface area for the absorption of phosphopeptides, while β-CD also offered a greater opportunity for the interaction between phosphopeptides and Ti 4+ . COOH-CNTs-CD-Ti with the aforementioned properities exhibited higher selectivity for phosphopeptides from the standard protein digests, the tryptic digests of nonfat milk and human serum, showing a great selective enrichment capability towards complex biological samples.
研究领域:
生物化学
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纳米生物学
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蛋白质组学
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PubMed
高影响因子
TALANTA
·
2017
·
IF 4.162
Jiebing Jiang,Xueni Sun,Yan Li,Chunhui Deng,Gengli Duan
Metal ions differed greatly in affinity towards phosphopeptides, and thus it is essential to systematically compare the phosphopeptides enrichment ability of different metal ions usually used in the IMAC techniques. In this work, for the first time, eight metal ions, including Nb 5+ , Ti 4+ , Zr 4+ , Ga 3+ , Y 3+ , In 3+ , Ce 4+ , Fe 3+ , were immobilized on the polydopamine (PDA)-coated Fe 3 O 4 (denoted as Fe 3 O 4 @PDA-M n+ ), and systematically compared by the real biosamples, in addition to standard phosphopeptides. Fe 3 O 4 microspheres were synthesized via the solvothermal reaction, followed by self-polymerization of dopamine on the surface. Then through taking advantage of the hydroxyl and amino group of PDA, the eight metal ions were easily adhered to the surface of Fe 3 O 4 @PDA. After characterization, the resultant Fe 3 O 4 @PDA-M n+ microspheres were applied to phosphopeptides enrichment based on the binding affinity between metal ions and phosphopeptides. According to the results, different metal ions presented diverse phosphopeptides enrichment efficiency in terms of selectivity, sensitivity and the enrichment ability from real complex samples, and Fe 3 O 4 @PDA-Nb 5+ and Fe 3 O 4 @PDA-Ti 4+ showed obvious advantages of the phosphopeptides enrichment effect after the comparison. This systematic comparison may provide certain reference for the use and development of IMAC materials in the future.
研究领域:
生物化学
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分析化学
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高影响因子
MOLECULES
·
2024
·Vol. 29
·期 7
·1656
·
IF 4.2
Qian Zhang,Linlin Hu,Jianyu Yang,Pengfei Guo,Jinhong Wang,Weifen Zhang
Immobilized metal ion affinity chromatography (IMAC) adsorbents generally have excellent affinity for histidine-rich proteins. However, the leaching of metal ions from the adsorbent usually affects its adsorption performance, which greatly affects the reusable performance of the adsorbent, resulting in many limitations in practical applications. Herein, a novel IMAC adsorbent, i.e., Cu(II)-loaded polydopamine-coated urchin-like titanate microspheres (Cu-PDA-UTMS), was prepared via metal coordination to make Cu ions uniformly decorate polydopamine-coated titanate microspheres. The as-synthesized microspheres exhibit an urchin-like structure, providing more binding sites for hemoglobin. Cu-PDA-UTMS exhibit favorable selectivity for hemoglobin adsorption and have a desirable adsorption capacity towards hemoglobin up to 2704.6 mg g−1. Using 0.1% CTAB as eluent, the adsorbed hemoglobin was easily eluted with a recovery rate of 86.8%. In addition, Cu-PDA-UTMS shows good reusability up to six cycles. In the end, the adsorption properties by Cu-PDA-UTMS towards hemoglobin from human blood samples were analyzed by SDS-PAGE. The results showed that Cu-PDA-UTMS are a high-performance IMAC adsorbent for hemoglobin separation, which provides a new method for the effective separation and purification of hemoglobin from complex biological samples.
研究领域:
生物化学
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材料科学
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生物技术
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PubMed
高影响因子
ANALYTICA CHIMICA ACTA
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2017
·
IF 4.95
Senwu Li,Kaiguang Yang,Lukuan Liu,Baofeng Zhao,Yuanbo Chen,Xiao Li,Lihua Zhang,Yukui Zhang
Tailor-made materials for the purification of proteins with His-tag was designed through synergizing the selectivity of surface sieving and metal ion affinity. By excluding impurity proteins out of the surface polymer network , such materials could purify His-tagged proteins from the crude cell lysis with purity up to 90%, improved by 14% compared to that obtained by the commercial metal chelating affinity materials. This study might promote the His-tagged protein purification to a new level.
研究领域:
生物化学
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蛋白质工程
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高影响因子
Analytical Methods
·
2018
·Vol. 10
·期 10
·1190-1195
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IF 2.073
Yinghua Yan,Yujie Lu,Mengying Chen,Hongze Liang
Immobilized metal affinity chromatography is a powerful method for phosphopeptide enrichment. However, the achievement of highly specific enrichment and sensitive detection of phosphopeptides by this method remains a big challenge due to the lack of high specificity and large binding capacity of conventional materials. In this work, adenosine tri-phosphate modified hydrophilic Fe3O4@PDA core–shell microspheres with Ti4+ exterior walls were successfully synthesized through a facile reaction at room temperature. The thicker grafting layer of the dopamine decoration exhibited enhanced hydrophilicity and biocompatibility, and the adenosine tri-phosphate molecule could offer superiorly strong and active metal phosphonate sites to bind phosphopeptides. The prepared microspheres demonstrated a low limit of detection (2 fmol), along with an exceptionally great specificity to capture phosphopeptides from a tryptic digest of the mixture of a nonphosphorylated protein BSA and a phosphorylated protein β-casein with molar ratios of BSA/β-casein up to 1000 : 1, and phosphopeptides from human serum and defatted milk were captured with high sensitivity and selectivity. In addition, the high magnetic susceptibility allowed convenient separation of the target peptides by magnetic separation. These outstanding features give the Ti4+-adenosine tri-phosphate-Fe3O4@polydopamine microspheres many advantages for mass spectrometric analysis of phosphopeptides.
研究领域:
生物化学
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蛋白质组学
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分析化学
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DOI
高影响因子
JOURNAL OF SEPARATION SCIENCE
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2010
·Vol. 33
·期 12
·1806-1815
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IF 2.551
Jian-Hong Wu,Yong Zhao,Ting Li,Cong Xu,Kuang Xiao,Yu-Qi Feng,Lin Guo
In our current work, we describe how open tubular-immobilized metal-ion affinity chromatography (OT-IMAC) capillary columns connected to a solid phase microextraction (in-tube SPME) device can be used for the enrichment of phosphopeptides. A phosphonate modified silica nanoparticle (NP)-deposited capillary was prepared by liquid phase deposition (LPD), and used for the immobilization of Fe 3+ , Zr 4+ or Ti 4+ . The enrichment capacities of three different OT-IMAC capillary columns were compared by using tryptically digested α-casein as sample. The improved extraction efficiency in our technique was demonstrated by comparing to a directly modified capillary, and a comparison of phosphopeptide extraction from simple and complex samples was tested for both modes. Our results show that the NP-IMAC-Zr 4+ capillary column can be used to selectively isolate phosphopeptides from real samples, and can enrich for β-casein phosphopeptides from concentrations as low as 1.7×10 −9 M.
研究领域:
分析化学
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蛋白质组学
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PubMed
高影响因子
Analytical Methods
·
2024
·Vol. 16
·期 5
·695-703
·
IF 2.7
Jiayi Wang,Xiaoya Zhang,Yinghua Yan,Rongrong Xuan
The comprehensive investigation of protein phosphorylation and glycosylation aids in the discovery of novel biomarkers as well as the understanding of the pathophysiology of illness. In this work, a nitrogen/titanium-rich porous organic polymer was developed by copolymerizing carbohydrazide (CH) and 2,3-dihydroxyterephthalaldehyde (2,3-Dha) and modifying with Ti4+ (CH–Dha–Ti4+). The adequate nitrogen contributes to the enrichment of glycopeptides via HILIC, while titanium benefits from capturing phosphopeptides through IMAC. The proposed method exhibits excellent selectivity (1 : 1000, both for glycopeptides and phosphopeptides), LOD (for glycopeptides: 0.05 fmol μL−1, for phosphopeptides: 0.2 fmol), loading capacity (for glycopeptides: 100 mg g−1, for phosphopeptides: 125 mg g−1) and size-exclusion effect (1 : 10 000, both for glycopeptides and phosphopeptides). Furthermore, CH–Dha–Ti4+ was applied to capture glycopeptides and phosphopeptides from human serum; 205 glycopeptides and 45 phosphopeptides were detected in the serum of normal controls; and 294 glycopeptides and 63 phosphopeptides were found in the serum of uremia patients after being analyzed by nano LC-MS/MS. The discovered glycopeptides and phosphopeptides were involved in several molecular biological processes and activities, according to a gene ontology study.
研究领域:
蛋白质组学
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生物化学
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分析化学
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DOI
PubMed
高影响因子
TALANTA
·
2017
·
IF 4.162
Heping Wang,Fenglong Jiao,Fangyuan Gao,Yayao Lv,Qiong Wu,Yan Zhao,Yehua Shen,Yangjun Zhang,Xiaohong Qian
To date, plenty of new alternative materials for phosphopeptides enrichment prior to mass spectrometry (MS) analysis appear, especially immobilized metal ion affinity chromatography (IMAC) materials. The variable combinations with different metal ions , chelating ligands and solid supports offer full of optionality for IMAC. However, further improvement was predicted by the tedious and complex synthetic process. In this work, a novel covalent organic framework (COF)-based IMAC material (denoted TpPa-2-Ti 4+ ) was prepared simply by direct immobilizing Ti (IV) into TpPa-2 COFs without any extra chelating ligands. The structure and composition of as-prepared composites were confirmed by PXRD, FT-IR and XPS, and a new flower-shaped Ti 4+ -IMAC with regular micro-nano hierarchical structure was observed in the SEM and TEM images. The obtained titanium (IV) ion-modified covalent organic frameworks demonstrated low limit of detection (4 fmol) and largely-satisfactory selectivity (β-casein: BSA=1:100) for phosphopeptide capturing from β-casein. Similarly, 18 and 17 phosphopeptides could be easily detected in the tryptic digest of α-casein or the digest mixture of α-casein and BSA (1:50). They were also successfully applied for enrichment of phosphopeptides from non-fat milk and HeLa cells with high sensitivity and satisfactory selectivity. All above results showed that the new titanium (IV) ion-modified covalent organic framework is expected to be a potential IMAC for phosphopeptide enrichment in large-scale phosphoproteomics studies.
研究领域:
生物化学
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蛋白质组学
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材料科学
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高影响因子
ANALYST
·
2015
·Vol. 140
·期 12
·4197-4205
·
IF 4.107
Caiyun Fang,Lei Zhang,Xiaoqin Zhang,Haojie Lu
Metal binding proteins play many important roles in a broad range of biological processes. Characterization of metal binding proteins is important for understanding their structure and biological functions, thus leading to a clear understanding of metal associated diseases. The present study is the first to investigate the effectiveness of magnetic microspheres functionalized with metal cations (Ca2+, Cu2+, Zn2+ and Fe3+) as the absorbent matrix in IMAC technology to enrich metal containing/binding proteins. The putative metal binding proteins in rat liver were then globally characterized by using this strategy which is very easy to handle and can capture a number of metal binding proteins effectively. In total, 185 putative metal binding proteins were identified from rat liver including some known less abundant and membrane-bound metal binding proteins such as Plcg1, Acsl5, etc. The identified proteins are involved in many important processes including binding, catalytic activity, translation elongation factor activity, electron carrier activity, and so on.
研究领域:
生物化学
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蛋白质组学
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医学
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PubMed
高影响因子
Journal of Materials Chemistry B
·
2018
·Vol. 6
·期 23
·3969-3978
·
IF 4.776
Bin Luo,Xiaoxi Zhou,Peipei Jiang,Qiangying Yi,Fang Lan,Yao Wu
Highly effective enrichment of phosphorylated biomolecules is vital for an in depth and comprehensive phosphoproteome analysis. Among affinity materials, materials based on the immobilized metal affinity chromatography (IMAC) method are most commonly used in phosphopeptide enrichment. However, the problem of metal cation loss in IMAC materials is still a challenge. In this study, a novel strategy has been developed for both highly effective enrichment of phosphorylated biomolecules and prevention of metal cation loss using Fe3O4/polydopamine/poly(2-aminoethyl methacrylate hydrochloride)/arginine (Fe3O4/PDA/PAMA–Arg) composite nanospheres. The magnetic composite nanospheres were prepared by a facile method, possessing uniform morphology, a high saturation magnetization (45 emu g−1), and a quick magnetic response (within 10 s). Noticeably, abundant guanidyl and amino groups of the PAMA–Arg brushes endowed the nanospheres with extremely high detection sensitivity (0.2 fmol), excellent selectivity (β-casein/BSA = 1 : 500), and high recyclability (5 cycles) in phosphopeptide analysis. Furthermore, the Fe3O4/PDA/PAMA–Arg nanospheres exhibited specific separation capacity towards β-casein or ovalbumin (OVA) in the model protein mixture and remarkably selective enrichment performance for phosphorylated biomolecules in real biological samples (egg white, nonfat milk, and rat brain lysate), showing great potential for trace biological detection and proteomics analysis.
研究领域:
分析化学
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生物化学
·
蛋白质组学
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DOI
PubMed
高影响因子
ANALYTICAL AND BIOANALYTICAL CHEMISTRY
·
2022
·Vol. 414
·期 27
·7885-7895
·
IF 4.478
Zhang Xiaoya,Feng Quanshou,Xie Zehu,Xu Fuxing,Yan Yinghua,Ding Chuanfan
In this work, on the basis of an immobilized metal ion affinity chromatography enrichment strategy, a new kind of covalent organic framework (COF) material for enrichment of phosphorylated peptides and exosomes was successfully prepared in a facile method, and Ti 4+ and Nb 5+ were used as dual-functional ions (denoted as COF-S–S-COOH-Ti 4+ /Nb 5+ ). With the advantage of unbiased enrichment towards phosphopeptides, COF-S–S-COOH-Ti 4+ /Nb 5+ shows ultra-high selectivity (maximum molar ratio of β -casein: BSA is 1:20,000) and low limit of detection (0.2 fmol). In addition, the material has an excellent phosphopeptide loading capacity (100 μg/mg) and reusability (at least seven times). Furthermore, applying the material to the actual sample, 4 phosphopeptides were selectively extracted from the serum of renal carcinoma patients. At the same time, exosomes with an intact structure in the serum of renal carcinoma patients were successfully isolated rapidly using this strategy. All experiments have shown that COF-S–S-COOH-Ti 4+ /Nb 5+ exhibits exciting potential in practical applications. Graphical abstract
研究领域:
分析化学
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生物医学工程
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PubMed
高影响因子
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