LWT-FOOD SCIENCE AND TECHNOLOGY
·
2024
·
IF 6
Yun Zhang,Yibing Zhang,Ruihua Lun,Qingshan Fu,Yuqiao Chang,Jiansen Du,Yi Zhang,Junping Yu
Conventional methods for Staphylococcus aureus ( S. aureus ) detection using mammalian immunoglobulins could yield false positive results, owing to the presence of the protein G-producing Streptococcus . In this study, because canine IgM can bind to protein A but not to protein G, we firstly designed a colorimetric platform based on canine IgM to identify S. aureus . Magnetic beads (MBs) functionalized with canine IgM acted as captors to enrich S. aureus , relying on the binding interaction between the Fc region of canine IgM and protein A in the bacterial cell wall, while horseradish peroxidase (HRP)-labeled rabbit anti- S. aureus IgG served as a tracer in the sandwich-type immunoassay, catalyzing the colorimetric reaction. Under optimal conditions, the proposed strategy achieved a satisfactory analytical performance with a broad linear range from 1.6 × 10 3 CFU/mL to 1.0 × 10 5 CFU/mL, a colorimetric minimum resolution of as low as 1.4 × 10 2 CFU/mL in 100 μL phosphate-buffered saline (PBS), and good selectivity in differentiating target from protein G-producing Streptococcus . Moreover, the proposed approach successfully identified S. aureus in various application scenarios (except for samples containing IgG), can be completed in less than 90 min, and was not dependent on any specialized equipment.
研究领域:
微生物学
·
免疫学
·
分析化学
✓ 已验证
产品关联
DOI
高影响因子
Surfaces and Interfaces
·
2025
·
IF 6.3
Dajun Zhang,Yufei Ma,Ying Wang,Xiangyu Liu,Mengfan Zhao,Yuanke Wang,Song Gao,Fu Ren,Qing Chen
A new solid-phase extraction agent, IgG@ Fe 3 O 4 -PEI, was successfully developed by conjugating immunoglobulin G (IgG) with amino-functionalized magnetic nanoparticles (Fe 3 O 4 -PEI). The IgG@Fe 3 O 4 -PEI composite was characterized using various techniques including infrared spectroscopy, Raman spectroscopy, magnetic property testing, Zeta potential measurements, energy spectrum analysis, scanning electron microscopy (SEM), and transmission electron microscopy (TEM). Because of the presence of a specific binding domain in the Fc fragment of IgG, the IgG@Fe 3 O 4 -PEI composite demonstrated highly selective adsorption of recombinant streptococcal G protein (rSPG) under optimal experimental conditions. At pH 6.0, the IgG@Fe 3 O 4 -PEI composite (0.5 mg) achieved a maximum adsorption efficiency of 93.96 % for rSPG at a concentration of 100 μg mL -1 . Adsorption model simulations indicated that the adsorption behavior followed the Langmuir adsorption model, with a theoretical maximum adsorption capacity of 125.0 mg g -1 . The enriched rSPG was eluted and recovered using Britton-Robinson (BR) buffer at pH 10.0, achieving a recovery efficiency of 95.8 %. In practical applications, the IgG@Fe 3 O 4 -PEI magnetic nanoparticles effectively enriched rSPG from an Escherichia coli expression system, and SDS-PAGE analysis confirmed the successful isolation of highly pure rSPG. Furthermore, the IgG@Fe 3 O 4 -PEI magnetic nanoparticles were used to capture proteins from human serum samples. After enrichment and processing, 430 proteins were identified, including 34 that were undetectable in untreated serum samples. This validated the high sensitivity and specificity of this method in serum proteomics, demonstrating its potential for isolating and identifying low-abundance proteins in complex biological samples. The developed system offers a valuable tool for advancing human serum proteomics research and targeted protein analysis.
研究领域:
生物化学
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纳米技术
·
蛋白质组学
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生物医学工程
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分子生物学
✓ 已验证
产品关联
DOI
高影响因子
INTERNATIONAL JOURNAL OF BIOLOGICAL MACROMOLECULES
·
2025
·
IF 8.5
Zhiqiang Guo,Mengdie Tian,Min Zhou,Zhuoer Li,Meijun Li
In this study, the formation of the immunoglobulin G-glabridin (IgG-GB) complex and nanocomplex was analyzed by assessing the structure, stability, solubility, and anticancer effects against the human epithelial ovarian cancer cell line SKOV3. The hydrodynamic sizes of the prepared IgG-GB nanocomplex were 190.1 ± 25.68 nm (PDI: 0.19 ± 0.02), and the solubility levels of GB in the free and nanoformulated forms were 97.28 μg/mL and 368.95 ± 59.63 μg/mL, respectively. Additionally, the IgG-GB nanocomplex exhibited a slower and more sustained release of GB in a pH-sensitive manner. Spectroscopic measurements indicated logKb values of 4.01 and 4.66 for the IgG-GB complex and the IgG-GB nanocomplex, respectively. Furthermore, the main reaction type between IgG and GB was found to be hydrophobic forces, which led to the partial folding of IgG and nanoformulated IgG. Cellular assays demonstrated that the IC50 concentrations of GB, the IgG-GB complex, and the IgG-GB nanocomplex in ovarian cancer SKOV3 cells were 10.74, 10.05, and 6.39 μM, respectively, while these amounts were > 77.84 μM in normal epithelial FHC cells. Moreover, the IgG-GB nanocomplex resulted in increased membrane leakage, mitochondrial dysfunction, and upregulation of Bax/Bcl-2 and caspase-3, along with caspase-3 activity determined by LDH, MP, real-time PCR, and enzyme activity assays. In conclusion, an efficient IgG-GB nanocomplex was potentially prepared and developed as a selective anticancer system against ovarian cancer cells.
研究领域:
药理学
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肿瘤学
·
生物化学
✓ 已验证
DOI
PubMed
高影响因子
JOURNAL OF CHROMATOGRAPHY B-ANALYTICAL TECHNOLOGIES IN THE BIOMEDICAL AND LIFE SCIENCES
·
2013
·
IF 2.487
Qiang Wu,Dong-Qiang Lin,Shan-Jing Yao
The aqueous two-phase system (ATPS) with polyethylene glycol (PEG) and hydroxypropyl starch (HPS) was evaluated for the extraction of human immunoglobulin G (IgG) from human serum albumin (HSA). The partitions of IgG in PEG/sulphate, PEG/phosphate, PEG/Dextran and PEG/HPS ATPSs were compared, and the results indicated that PEG/HPS system was most suitable for IgG extraction. The effects of the concentrations of PEG, HPS and NaCl addition and pH on the partition of IgG and HSA in PEG/HPS ATPS were investigated. It was found that with 15% NaCl addition at pH 8.0 IgG could be largely recovered in the top PEG-rich phase and most of HSA kept in the bottom phase. In addition, the response surface methodology (RSM) was used to optimize the extraction of IgG from the protein mixture of IgG and HSA. The optimal conditions were obtained as 12% (w/w) PEG 4000, 18% (w/w) HPS and 10% (w/w) NaCl at pH 8.0. The extraction yield of IgG in the top phase was 99.2% and the purification factor could reach 5.28. The back extractions of IgG into a phosphate-rich bottom phase were also studied. The total purification factor was 5.73 with the yield of 84.0%. The results indicated that PEG/HPS ATPS might be a promising alternative for the primary recovery of IgG from the complicated feedstock.
研究领域:
生物化学
·
蛋白质工程
✓ 已验证
DOI
高影响因子
JOURNAL OF CHEMICAL AND ENGINEERING DATA
·
2019
·
IF 2.298
Hong-Yun Zhu,Sheng-Gang Chen,Qilei Zhang,Dong-Qiang Lin,Shan-Jing Yao
Alternative antibody-binding affinity ligands are expected to support robust time- and cost-effective antibody purification with rapid improvement of upstream productivity and demand on downstream process optimization. Five new tetrapeptide biomimetic affinity ligands (FYRH, HWRH, YHRI, HYRF, and FHRA) were screened by modeling the molecular interaction mechanism between tetrapeptide ligands and the Fc fragment of IgG with molecular dynamics simulation and the flexible docking method, and the corresponding five resins were prepared in this work to evaluate their adsorption characteristics of human immunoglobulin G (hIgG). The results showed that the binding capacities to hIgG of the five resins showed a strong pH-dependency, which was high at pH 7.0–9.0, and extremely low at pH 5.0–6.0, providing a benefit choice to separate hIgG. The dynamic binding capacities (DBC) at 10% breakthrough of five resins could reach up to 28–37 mg/mLgel, and Ac-FYRH-4FF resin showed the best adsorption performance. The elution test results with Ac-FYRH-4FF resin indicated that hIgG could be eluted mildly at pH 5.0 with a 96.5% recovery, meaning that this resin can be considered as a powerful candidate of alternative biomimetic affinity chromatographic resin.
研究领域:
生物化学
·
蛋白质纯化
✓ 已验证
产品关联
DOI
高影响因子
BIOCHEMICAL ENGINEERING JOURNAL
·
2016
·
IF 2.463
Rong-Zhu Wang,Dong-Qiang Lin,Wen-Ning Chu,Qi-Lei Zhang,Shan-Jing Yao
Peptide ligands have been developed as powerful alternatives to Protein A for antibody purification. In this study, a tetrapeptide library with two aromatic residues, one arginine and one aromatic/aliphatic residue (Aro-Aro-R-Aro/Ali) was constructed considering molecular interactions and applications. The library had 128 peptide ligands and was screened by flexible docking and molecular dynamics simulation. The results indicated that Ac-YFRH was the best ligand, and then was coupled onto an aminated argarose matrix. The results of isotherm adsorption showed that the binding capacity of human immunoglobulin G (IgG) was high at pH 7⿿8 and low at pH 4⿿5. The IgG adsorption also showed obvious salt-independence, and the addition of salt could reduce unspecific adsorption caused by electrostatic interaction. After the optimization of pH and salt addition, high IgG purity of 98.4% could be obtained with the recovery of 89.4% from BSA-containing feedstock. The purification of monoclonal antibody from CHO cell culture supernatant was tested, and the purity and recovery reached 98.0% and 79.5%, respectively. The results demonstrated that the new resin with the tetrapeptide YFRH ligand had high affinity and flexible operation window for IgG purification.
研究领域:
生物化学
·
分子生物学
✓ 已验证
产品关联
DOI
高影响因子
ACS Applied Materials & Interfaces
·
2019
·
IF 8.456
Quan-Quan Zhuang,Hao-Hua Deng,Shao-Bin He,Hua-Ping Peng,Zhen Lin,Xing-Hua Xia,Wei Chen
Few-atom gold nanoclusters (AuNCs) have been fabricated and used for various fields owing to their remarkable optical and photophysical features. However, the rational design for the antibody-mediated synthesis of fluorescent AuNCs for direct antigen–antibody reactions remains unexplored. In this work, immunoglobulin G (IgG)-functionalized AuNCs (IgG-AuNCs) were successfully prepared via a facile and fast biomineralization process. The generated IgG-AuNCs can emit intense red fluorescence with a high photoluminescence quantum yield. Besides strong emission, the bioactivity of IgG on the IgG-AuNCs can be retained. Surface plasmon resonance measurements suggested that IgG-AuNCs can bind to goat anti-human IgG with an affinity constant of 6.21 × 10–8 M. A simple detection method was then developed using a dot-blot immunoassay with IgG-AuNCs as fluorescent tags. Experimental results confirmed that the IgG-AuNC-based fluorescent reporters had many advantages such as low nonspecific adsorption and good photostability, offering immense potential for the development of efficient biosensors. This work can be extended to other specific antibodies to produce multifunctional AuNCs and utilized to detect and monitor targeted analytes and biological events of interest.
研究领域:
生物化学
·
纳米生物学
·
分析化学
✓ 已验证
产品关联
DOI
PubMed
高影响因子
JOURNAL OF CHROMATOGRAPHY B-ANALYTICAL TECHNOLOGIES IN THE BIOMEDICAL AND LIFE SCIENCES
·
2019
·
IF 2.813
Jiawen Li,Dongqiang Lin,Shanjing Yao,N.K.H. Slater,Qilei Zhang
Traditional chromatographic techniques used in downstream processes of biomolecule manufacturing are often time-consuming and expensive. In this study, a cost-effective microporous micro-capillary film (MMCF) composed of ethylenevinyl alcohol (EVOH) was evaluated for its potential application in immunoadsorption with high process efficiency. A peptide ligand Ac-Phe-Tyr-His-Glu (Ac-FYHE) was immobilized on the inner surface of MMCF for selective binding of human immunoglobulin (hIgG). The porous structure and chemical properties of the prepared MMCF were confirmed by scanning electron microscopy (SEM) and Fourier transform infrared spectroscopy (FTIR). hIgG (2 mg/ml) adsorption studies demonstrated that the binding process followed a Langmuir isotherm with equilibrium adsorption capacities of 9.31 and 3.47 mg/ml adsorbent under static and dynamic conditions, respectively. Moreover, the membrane showed good flowrate tolerance when studied under flowrates of 0.5 ml/min to 10 ml/min. hIgG purity was 88.2% when obtained from an hIgG (2 mg/ml) and HSA (8 mg/ml) mixture and the purity remained over 80.0% when hIgG concentrations increased in the mixtures. Moreover, purity of 82.3% was achieved when removing hIgG directly from human serum. The MMCF-Ac-FYHE affinity column is expected to selectively remove hIgG from blood for the treatment of autoimmune diseases with high efficiency and cost effectiveness.
研究领域:
生物工程
·
医学工程
✓ 已验证
产品关联
DOI
PubMed
高影响因子
JOURNAL OF SEPARATION SCIENCE
·
2022
·Vol. 45
·期 15
·2946-2955
·
IF 3.614
Mengting Li,Dongqiang Lin,Shanjing Yao,Qilei Zhang
Molecular interactions between ligands and target biomolecules are crucial in the development of chromatographic techniques for the separation and purification of biotherapeutics. In this study, the role of functional moieties on a mixed-mode ligand (phenylalanine-tyrosine-glutamate-5-aminobenzimidazole) for human immunoglobulin G purification was investigated and a detailed mechanism was discussed. A similar ligand with glutamic acid substituted by glutamine (phenylalanine-tyrosine-glutamine-5-aminobenzimidazole) together with other resins including a commercial resin (CM Bestarose Fast Flow), phenylalanine-tyrosine-glutamate, glutamate-5-aminobenzimidazole, and 5-aminobenzimidazole resins were prepared for comparison. Molecular dynamics simulation and experimental studies were used to analyze the difference between these ligands. The results showed that the carboxyl group of phenylalanine-tyrosine-glutamate-5-aminobenzimidazole contributed 70% of the electrostatic interaction during human immunoglobulin G binding, and 5-aminobenzimidazole provided electrostatic repulsion for desorption, which showed low selectivity and binding capacities at pH 4.0 (dynamic binding capacities at 10% breakthrough of human immunoglobulin G = 1.0 mg/ml resin, dynamic binding capacities at 10% breakthrough of human serum albumin = 1.2 mg/ml resin) when used as an individual resin ligand. The results showed in this study demonstrated that it is possible to achieve optimal antibody separation and purification through reasonable ligand design by understanding the performance of key functional moieties in binding and elution processes.
研究领域:
生物化学
·
生物分离技术
✓ 已验证
产品关联
DOI
PubMed
高影响因子
Microsystems & Nanoengineering
·
2024
·Vol. 10
·期 1
·1-10
·
IF 7.3
Zhou Bin,Hu Chao,Li Haoyang,Ye Xiangyi,Wen Baohua,Zhou Zhangkai,Cai Jingxuan,Zhou Jianhua
Surface lattice resonances (SLR) have been demonstrated to enhance the sensitivity and reduce the full width at half maximum (FWHM) of the plasmonic resonances. However, their widespread application in immunoassays has been hindered by limitations of high structural defect sensitivity and fabrication costs. Here, we design a novel three-layer cylindrical SLR array that exhibits high tolerance against structural defects, which would facilitate straightforward fabrication. By integrating metal evaporation and nanoimprint lithography, we demonstrate the replication of the SLR array with exceptional quality. Theoretical simulations indicate that the resonance dips of these arrays exhibit are not sensitive to various structural defects. The experimental results reveal that the FWHM of these arrays can be as low as 5.1 nm while maintaining robust resonance characteristics. Furthermore, we demonstrated the high spectral sensitivity of the SLR array, which enabled the detection of immunoglobulin G (IgG) at concentrations as low as 609 pg/mL. These findings emphasize the potential of the defect-insensitive SLR array as a highly scalable immunoassay platform with exceptional performance.
研究领域:
生物医学工程
·
光学传感
·
纳米技术
✓ 已验证
产品关联
DOI
PubMed
高影响因子
ACS Sensors
·
2018
·
IF 5.711
Nianzu Liu,Ni Hui,Jason J. Davis,Xiliang Luo
The construction of sensitive and selective biosensors capable of detecting specific targets in complex biological samples remains a challenge highly relevant to a range of sensor/diagnostic applications. Herein, we have utilized a multifunctional peptide to present an interface that supports the very specific recruitment of targets from serum. The novel peptide sequence designed contains an anchoring domain (CPPPP-), an antifouling domain (-NQNQNQNQDHWRGWVA), and a human immunoglobulin G (IgG) recognition domain (-HWRGWVA), and the whole peptide was designed to be antifouling. These were integrated into polyaniline nanowire arrays in supporting the quantification of IgG (with a limit of detection of 0.26 ng mL–1) in neat serum and real clinical samples. The strategy of utilizing multisegment peptide films to underpin highly selective target recruitment is, of course, readily extended to a broad range of targets for which an affinity sequence can be generated.
研究领域:
生物化学
·
诊断学
·
材料科学
✓ 已验证
DOI
PubMed
高影响因子
Cancers
·
2022
·Vol. 14
·期 5
·1333
·
IF 6.575
Jichen Sha,Rongrong Zhang,Jiteng Fan,Yong Gu,Yiqing Pan,Jing Han,Xiaoyan Xu,Shifang Ren,Jianxin Gu
Simple SummaryAs serum IgG glycosylation is associated with various cancers, our goal is to explore whether serum IgG galactosylation and its associated glycans could be used as tumor markers associated with hepatocellular carcinoma (HCC). At the same time, we explore the effect of the B-cell-specific ablation of B4GALT1 on HCC and finally analyze whether the low incidence of female cancer was related to the findings from the above perspective. The results demonstrate that the tumor marker of serum IgG glycosylation is galactosylation and its associated glycans and that the B-cell-specific ablation of B4GALT1 reduces HCC formation by reducing serum IgG galactosylation levels and by modulating the associated glycans, meaning that the lower incidence of cancer in women may be related to minor changes in the B-cell B4GALT1 and unchanged serum IgG galactosylation levels. This study aims to provide a theoretical basis for the early diagnosis and prevention of HCC and to determine why it has such a high incidence in males.AbstractSerum immunoglobulin G (IgG) glycosylation, especially galactosylation, has been found to be related to a variety of tumors, including hepatocellular carcinoma (HCC). However, whether IgG glycan changes occur in the early stages of HCC formation remains unclear. We found that the galactosylation level increased and that the related individual glycans showed regular changes over the course of HCC induction. Then, the effect of the B-cell-specific ablation of β1,4galactosyltransferase 1 (CKO B4GALT1) and B4GALT1 defects on the IgG glycans that were modified during the model induction process and HCC formation is investigated in this study. CKO B4GALT1 reduces serum IgG galactosylation levels and reduces cancer formation. Furthermore, insignificant changes in the B-cell B4GALT1 and unchanged serum IgG galactosylation levels were found during cancer induction in female mice, which might contribute to the lower cancer incidence in female mice than in male mice. The gender differences observed during glycan and B4GALT1 modification also add more evidence that the B4GALT1 in B cells and in serum IgG galactosylation may play an important role in HCC. Therefore, the findings of the present research can be used to determine the methods for the early detection of HCC as well as for prevention.Keywords:hepatocellular carcinoma;immunoglobulin G;glycan;β1,4-galactosyltransferase 1;mouse
研究领域:
医学
·
分子生物学
✓ 已验证
DOI
PubMed
高影响因子
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