TALANTA
·
2018
·
IF 4.244
Dandan Jiang,Xiqian Li,Xueju Lv,Qiong Jia
Dendrimers exhibit tunable terminal functionality and bio-friendly nature, making them of being promising materials for applications in the field of separation and enrichment. In this work, we prepared magnetic hydrazide-functionalized poly-amidoamine (PAMAM) dendrimer embedded with TiO 2 for the enrichment of phosphopeptides. The novel affinity probe possessed superparamagnetism , realizing its rapid separation from sample solution. Electrostatic attraction and hydrogen bonding existed between PAMAM and phosphopeptides while Lewis acid-base interaction was originated between TiO 2 and the targets. The combined synergistic strength of multiple binding interactions contributed to the highly selective enrichment of phosphopeptides. The specificity for the capture of phosphopeptides was reflected in quantities as low as 1:1000 mass ratio of phosphopeptides to non-phosphopeptides. The detection limit of β -casein digests was low to 0.4 fmol, indicating the high sensitivity of the developed method. Fifteen and four phosphopeptides could be selectively captured from non-fat milk digests and human serum samples, which further confirmed the great potential of the affinity probe in the extraction of low-abundance phosphopeptides from real complex biological samples.
研究领域:
生物化学
·
分析化学
✓ 已验证
产品关联
DOI
PubMed
高影响因子
JOURNAL OF NANOBIOTECHNOLOGY
·
2019
·Vol. 17
·期 1
·1-11
·
IF 5.345
Hou Zhanwu,Wang Zhen,Liu Run,Li Hua,Zhang Zhengyi,Su Tian,Yang Jeffy,Liu Huadong
Background Gold nanoparticles (AuNPs) have been proposed for many applications in medicine and bioanalysis. For use in all these applications, maintaining the stability of AuNPs in solution by suppressing aggregation is paramount. Herein, the effects of amino acids were investigated in stabilizing AuNPs by rationally designed peptide scaffolds.Results Compared to other tested amino acids, phosphotyrosine (pY) significantly stabilized AuNPs. Our results indicated that pY modified AuNPs presented a high level of stability in various solutions, and had good biocompatibility. When a pY-peptide was used in stabilizing AuNPs, the phosphate group could be removed by phosphatases, which subsequently caused the aggregation and the cargo release of AuNPs. In vitro study showed that AuNPs formed aggregation in a phosphatase concentration depending manner. The aggregation of AuNPs was well correlated with the enzymatic activity (R2 = 0.994). In many types of cancer, a significant increase in phosphatases has been observed. Herein, we demonstrated that cancer cells treated with pY modified AuNPs in conjunction with doxorubicin killed SGC-7901 cells with high efficiency, indicating that the pY peptide stabilized AuNPs could be used as carriers for targeted drug delivery.Conclusion In summary, pY peptides can act to stabilize AuNPs in various solutions. In addition, the aggregation of pY-AuNPs could be tuned by phosphatase. These results provide a basis for pY-AuNPs acting as potential drug carriers and anticancer efficacy.
研究领域:
生物化学
·
纳米医学
·
肿瘤学
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产品关联
DOI
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
·
分析化学
✓ 已验证
DOI
高影响因子
JOURNAL OF CHROMATOGRAPHY A
·
2017
·
IF 3.981
Xiao-Mei He,Xi Chen,Bi-Feng Yuan,Yu-Qi Feng
Grafting copolymerization, especially “grafting from” approach, has attracted a great attention for the preparation of cellulose-based materials with various functionalities. In this study, a novel phosphate group-containing cotton fiber-based material, CF-NH 2 -AZO-p(VPA-x), was successfully synthesized by a “grafting from” radical polymerization approach using vinyl phosphonic acid (VPA) as monomer. The phosphate group content of CF-NH 2 -AZO-p(VPA-x) could be easily regulated by adjusting the concentration of VPA monomer. Subsequently, titanium ions immobilized CF-NH 2 -AZO-p(VPA-x) (CF-NH 2 -AZO-p(VPA-x)-Ti 4+ ) was prepared and used as an immobilized metal affinity chromatography (IMAC) adsorbent for the selective enrichment of phosphopeptides from various biological samples. The relationship between enrichment efficiency and phosphate group content of CF-NH 2 -AZO-p(VPA-x)-Ti 4+ was investigated. The optimized CF-NH 2 -AZO-p(VPA-2)-Ti 4+ (x = 2) exhibited high selectivity and extraction capacity in phosphopeptides enrichment from standard peptides mixture, non-fat milk digests and protein-rich human serum. In addition, CF-NH 2 -AZO-p(VPA-2)-Ti 4+ was further applied for the specific capture of phosphopeptides from tryptic digests of rat brain lysate, and 3241 unique phosphopeptides were identified from 0.5 mg of rat brain by combining CF-NH 2 -AZO-p(VPA-2)-Ti 4+ pretreatment with liquid chromatography–tandem mass spectrometry (LC–MS/MS) analysis. We expect the proposed method can promote the development of “grafting from” approach in the preparation of cellulose-based adsorbents and broaden the application of cellulose-based adsorbents for biological analysis.
研究领域:
生物化学
·
材料科学
✓ 已验证
DOI
PubMed
高影响因子
JOURNAL OF COLLOID AND INTERFACE SCIENCE
·
2013
·
IF 3.172
Zhi-Gang Wang,Gong Cheng,Yan-Lin Liu,Ji-Lin Zhang,De-Hui Sun,Jia-Zuan Ni
In this work, novel magnetic polymeric core–shell structured microspheres with immobilized Ce(IV), Fe 3 O 4 @SiO 2 @PVPA-Ce(IV), were designed rationally and synthesized successfully via a facile route for the first time. Magnetic Fe 3 O 4 @SiO 2 microspheres were first prepared by directly coating a thin layer of silica onto Fe 3 O 4 magnetic particles using a sol–gel method, a poly(vinylphosphonic acid) (PVPA) shell was then coated on the Fe 3 O 4 @SiO 2 microspheres to form Fe 3 O 4 @SiO 2 @PVPA microspheres through a radical polymerization reaction, and finally Ce(IV) ions were robustly immobilized onto the Fe 3 O 4 @SiO 2 @PVPA microspheres through strong chelation between Ce(IV) ions and phosphate moieties in the PVPA. The applicability of the Fe 3 O 4 @SiO 2 @PVPA-Ce(IV) microspheres for selective enrichment and rapid separation of phosphopeptides from proteolytic digests of standard and real protein samples was investigated. The results demonstrated that the core–shell structured Fe 3 O 4 @SiO 2 @PVPA-Ce(IV) microspheres with abundant Ce(IV) affinity sites and excellent magnetic responsiveness can effectively purify phosphopeptides from complex biosamples for MS detection taking advantage of the rapid magnetic separation and the selective affinity between Ce(IV) ions and phosphate moieties of the phosphopeptides. Furthermore, they can be effectively recycled and show good reusability, and have better performance than commercial TiO 2 beads and homemade Fe 3 O 4 @PMAA-Ce(IV) microspheres. Thus the Fe 3 O 4 @SiO 2 @PVPA-Ce(IV) microspheres can benefit greatly the mass spectrometric qualitative analysis of phosphopeptides in phosphoproteome research.
研究领域:
分析化学
·
蛋白质组学
✓ 已验证
产品关联
DOI
PubMed
高影响因子
TALANTA
·
2017
·
IF 4.162
Xing-Yu Long,Jia-Yuan Li,Dong Sheng,Hong-Zhen Lian
The spinel-type magnetic manganese ferrite (MnFe 2 O 4 ) microspheres synthesized by simple solvothermal method were used as a novel adsorbent for selective enrichment and effective isolation of phosphopeptides. The uniform MnFe 2 O 4 magnetic affinity microspheres (MAMSs) had a narrow particle size distribution between 250 and 260 nm, and displayed superparamagnetism with a saturation magnetization value of 67.0 emu/g. Comprehensively, the possible formation mechanism of MnFe 2 O 4 microspheres with ferric and manganous sources as dual precursors was elucidated by comparison with those of Fe 3 O 4 nanoparticles and MnOOH nanosheets respectively with either ferric or manganous source as single precursor. It was suggested that the spherical or sheet nanostructures could be achieved via secondary recrystallization or Ostwald ripening . The MnFe 2 O 4 MAMSs probe exhibited excellent dispersibility in aqueous solution, and rapid magnetic separation within 15 s, as well as good reusability. More importantly, MnFe 2 O 4 was highly selective for phosphopeptides because of the strong coordination interaction between metal ions (Fe 3+ and Mn 2+ ) and phosphate groups of phosphopeptdies. This high specificity was demonstrated by effectively enriching phosphopeptides from digest mixture of β -casein and bovine serum albumin (BSA) with high content of non-phosphopeptides, and embodied further in phosphopeptides enrichment from non-fat milk digests and human serum. Consequently, the prepared MnFe 2 O 4 affinity materials are expected to possess great potential in phosphoproteome research.
研究领域:
生物化学
·
材料科学
·
蛋白质组学
✓ 已验证
产品关联
DOI
PubMed
高影响因子
Talanta Open
·
2025
·
IF 3.7
Dong Wei,Hao Zhang,Mingyang Li,Liuxi Chu
Circulating extracellular vehicles (EVs) play a crucial role in intercellular communication by transporting a diverse array of biological signaling molecules. Growing evidence indicates that phosphoproteins within these EVs hold promise as biomarkers for diagnostic and prognostic assessments, particularly in oncology. In this study, EVtrap (extracellular vesicle total recovery and purification) was utilized to isolate urinary EVs. We then combined this with an advanced liquid chromatography-tandem mass spectrometry (LC−MS/MS) technique to perform neutral loss-information-dependent acquisition-enhanced product ion (NL-IDA-EPI) analysis. This technique is specifically tailored to quantify endogenous phosphopeptides in urinary EVs by monitoring a consistent mass loss of 49 Da (H 3 PO 4 , doubly charged state) in the MS2 spectra. In this study, over 2,000 putative phosphopeptide signals were detected in a 10-mL urine sample without the need for prior phosphopeptide enrichment, thus simplifying the process and potentially minimizing sample bias. Comparative analysis of samples from high- and low-risk prostate cancer patients and a healthy control group revealed six differentially expressed phosphopeptides. These phosphopeptides serve as a promising panel of potential biomarkers, highlighting the effectiveness of our analytical framework in tracking cancer-related signaling events via phosphorylation within EVs using NL-IDA-EPI.
研究领域:
医学
·
蛋白质组学
·
肿瘤学
✓ 已验证
产品关联
DOI
高影响因子
LANGMUIR
·
2023
·
IF 3.9
Ruizhi Tang,Lei Pan,Quan Bai,Cong Li,Shujuan Ma,Junjie Ou,Yehua Shen
High-performance reusable materials from renewable resources are rare and urgently required in bioseparation. Herein, a series of tannic acid-chitosan composite membranes for the enrichment of phosphopeptides were fabricated by the freeze casting method. First, a tannic acid-chitosan composite membrane was acquired via the multiple hydrogen bonds between tannic acid and chitosan, which had a long-range aligned three-dimensional microstructure. Second, a covalent–hydrogen bond hybrid composite was also fabricated, with stable and aligned honeycomb-like microstructures that formed by the synergy of covalence and hydrogen bonding. Besides, a ternary composite membrane was “one-pot” synthesized by the copolymerization of tannic acid, chitosan, and Ti4+ ions, indicating the feasibility of involving metal ions in the composition of the polymer skeleton in place of additional modification steps. The as-prepared chitosan composite membranes exhibited excellent performance in the enrichment of phosphopeptides from β-casein tryptic digest and human serum. Benefitting from the long-range aligned honeycomb-like structure coordinated by hydrogen bonds and covalent bonds, and a large number of pyrogallol functional groups provided by tannic acid, the covalent–hydrogen bond hybrid membrane showed excellent reusability and could be reused up to 16 times in phosphopeptide enrichment, as far as we know, which is the best reported result to date.
研究领域:
生物分离
·
材料科学
·
生物化学
✓ 已验证
DOI
PubMed
高影响因子
Journal of Materials Chemistry B
·
2013
·Vol. 1
·期 10
·1491-1500
·
IF 0
Zhi-Gang Wang,Gong Cheng,Yan-Lin Liu,Ji-Lin Zhang,De-Hui Sun,Jia-Zuan Ni
The multifunctional microspheres consisting of the magnetic γ-Fe2O3 core and the affinity REVO4 (RE = Sm, Dy, Ho) shell have been synthesized via the homogenous precipitation–calcination–ion exchange three-step synthetic route. Their morphologies, structures, surface properties, and magnetisms were characterized, respectively. SEM and TEM images indicate that they all have an average size of about 400 nm and very rough surfaces. The TEM images further reveal that the γ-Fe2O3@REVO4 microspheres are all core–shell structures and the REVO4 shells are about 55–60 nm in thickness. The XRD pattern analyses show that the magnetic γ-Fe2O3 cores belong to cubic structure and the REVO4 (RE = Sm, Dy, Ho) shells are composed of their corresponding tetragonal major phases. HRTEM images, FTIR spectra and EDS further demonstrate the formations of tetragonal REVO4 shells based on checkup of the corresponding lattice fringes, characteristic IR absorption peaks and element signals. Their potentials for selective capture, rapid separation, and convenient mass spectra (MS) labeling of the phosphopeptides from complex proteolytic digests are explored and evaluated for the first time. The experimental results show that the magnetic γ-Fe2O3@REVO4 core–shell structured microspheres have high selective affinity for the phosphopeptides. The trapped phosphopeptides can be rapidly isolated by an external magnetic field, and can be easily identified by characteristic MS signals from 80 Da mass losses in the mass spectra (MS). Additionally, the γ-Fe2O3@REVO4 affinity materials can be reused after recovery.
研究领域:
材料科学
·
分析化学
·
生物化学
✓ 已验证
DOI
PubMed
LWT-FOOD SCIENCE AND TECHNOLOGY
·
2021
·
IF 4.952
Liangjie Tian,Dandan Xiong,Jie Jia,Xuebo Liu,Ya Zhang,Xiang Duan
Lactic acid bacteria fermentation is a safe and green technology that can modify the function of food ingredients (including proteins). In this study, phosvitin (PSV) was subjected to fermentation with lactic acid bacteria for 0, 3, 6 and 9 h, respectively. After fermentation treatment, PSV was partially degraded into short peptides such as K.KPMDEEENDQVK.Q and D.EEENDQVK.Q. The surface hydrophobicity, thermal stability and apparent viscosity were significantly improved. Correspondingly, the emulsifying activity of PSV was enhanced from 12.65 to 16.38, 16.81 and 14.98 m 2 /g after 3, 6 and 9 h of fermentation, while the emulsifying stability increased from 34.08 min to 44.18, 60.49 and 40.45 min, respectively. In parallel, the phosvitin phosphopeptides (PPP) derived from fermentation-treated PSV showed a higher calcium-binding capacity compared with native PSV. These results indicated that lactic acid bacteria fermentation has the potential as a strategy to improve the emulsifying properties and bioactivity of PSV.
研究领域:
食品科学
·
生物技术
·
蛋白质化学
✓ 已验证
DOI
高影响因子
JOURNAL OF CHROMATOGRAPHY B-ANALYTICAL TECHNOLOGIES IN THE BIOMEDICAL AND LIFE SCIENCES
·
2022
·
IF 3.318
Zong-Qin Ma,Ying-Hui Wang,Yuan Peng,Xin Guo,Zhe Meng
In this paper, we developed a facile route for the preparation of a novel bimetal oxide affinity chromatography (MOAC) material. The TiO 2 /ZrO 2 @MoS 2 was constructed by the electrostatic interaction between titanium oxide/zirconia ( w : w , 10:1) and molybdenum disulfide nanosheet. The nanocomposite has the large specific surface area (186.30 m 2 ⋅g −1 ) and pore volume (0.37 cm 3 ⋅g −1 ). Compared with single-metal probes, the combination of bimetallic oxides probe (TiO 2 /ZrO 2 ) and hydrophilicity MoS 2 support offered multitudinous affinity sites for phosphopeptides capturing from tryptic digests of protein samples under 50% acetonitrile-1% trifluoroacetate conditions. Singnificant feasibility of the TiO 2 /ZrO 2 @MoS 2 nanomaterial for the enrichment of phosphopeptides under optimal conditions was proved via the bovine serum albumin (BSA) and the mixtures of β-casein. The phosphopeptide expression was identified using ultra-performance liquid chromatography (uHPLC) separation and-linear ion trap mass spectrometry (MS n ). With these affinity characters of TiO 2 /ZrO 2 @MoS 2 , it exhibited higher binding capacity (25 mg⋅g −1 ), better selectivity for phosphopeptides from β-casein/BSA (1:2000) tryptic digests, high sensitivity (1 fmol⋅µL −1 ) towards phosphopeptides from β-casein tryptic digests, and great reusability of 8 cycles test for capturing phosphopeptides. In addition, the TiO 2 /ZrO 2 @MoS 2 with high sensitivity and selectivity was successfully applied to enriching phosphopeptides from nonfat milk and human serum samples. More importantly, the TiO 2 /ZrO 2 @MoS 2 was further successfully applied to multi-phosphopeptides enrichment, 1779 serine, threonine and tyrosine phosphosites can be identified in A549 cell protein tryptic digest. Compared with commercial TiO 2 from enrichening 416 phosphopeptide from A549 cell lysates, the successful locating of 44 phosphosites were overlapped.
研究领域:
生物化学
·
分析化学
·
蛋白质组学
✓ 已验证
产品关联
DOI
PubMed
高影响因子
Analytical Methods
·
2018
·Vol. 10
·期 10
·1190-1195
·
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
·
蛋白质组学
·
分析化学
✓ 已验证
DOI
高影响因子
危险品化学品经营许可证（带存储）