Food & Function
·
2023
·
IF 6.1
Xian Xia,Jiangdong Li,Rui Liang,Yi Li,Xiaojuan Ma,Ying Yang,Daniel Lozano-Ojalvo
Ovomucoid is the immune-dominant allergen in the egg white of hens. Due to its structure based on nine disulfide bonds as well as its resistance to heat and enzymatic hydrolysis, the allergenicity of this food protein is difficult to decrease by technological processes. We sought to reduce its allergenicity through the Maillard reaction. The unfolding of ovomucoid with L-cysteine-mediated reduction was used to increase accessibility to conformational and linear epitopes by modifying the secondary and tertiary structures of the allergen. Glycation with different saccharides revealed the beneficial effect of maltose glycation on the IgG-binding capacity reduction. By determining the better glycation conditions of unfolded ovomucoid, we produced ovomucoid with reduced IgE binding capacity due to the glycation sites (K17, K112, K129, and K164) on epitopes. Moreover, after simulated infant and adult gastrointestinal digestion, the unfolded plus glycated ovomucoid showed higher ABTS˙+ scavenging activity, O2˙− scavenging activity, ˙OH scavenging activity, Fe2+ chelating activity, and a FRAP value; in particular, for ˙OH scavenging activity, there was a sharp increase of more than 100%.
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CHINESE JOURNAL OF ANALYTICAL CHEMISTRY
·
2013
·
IF 0.769
Jian-Ming JI,Xiu-Xia HE,Qian DUAN,Zhen-Xin WANG
Uniformly porous silicon substrate with large surface area (12 mm × 58 mm) was prepared by double-cell electrochemical etching method. After functionalized with epoxide groups, the porous silicon substrate can be employed to fabricate protein microarray. In proof of principle experiment, well-known biomolecule recognition system (the interaction of mouse IgG with rabbit anti-mouse IgG antibody) was chosen to establish the new microarray format. It was found that the detection sensitivity of the mouse IgG microarray was dependent on preparing conditions of porous silicon substrate including etching current density, etching time, and concentration of HF. Under the optimal conditions (500 mA cm −2 etching current density, 450 s etching time and 25% HF), specific binding could be detected using rabbit anti-mouse IgG antibody at concentrations as low as 10 ng mL −1 with a dynamic range from 316 ng mL −1 to 10 μg mL −1 . The results demonstrate that the as-prepared porous silicon substrate could be used to fabricate the protein microarray with high reaction capability.
研究领域:
生物技术
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材料科学
·
分析化学
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BIOTECHNOLOGY AND BIOPROCESS ENGINEERING
·
2017
·Vol. 22
·期 1
·76-82
·
IF 1.16
Gu Jia-Li,Tong Hong-Fei,Sun Lai-Yu
Macroporous magnetic agarose particles (MMAPs) were prepared with calcium carbonate as the porogent by the water-in-oil suspension thermal regeneration method. MMAPs with good sphericity and appropriate particle size were obtained. The physical properties of the beads were determined and it was found that the water content (92.1%), porosity (94.4%) and mean pore diameter (120.1 nm) of the MMAPs were higher than those for the normal magnetic particles, indicating successful generation of macropores after calcium carbonate addition. Compared with normal magnetic particles, the mass transfer of biomolecules in MMAPs was remarkably enhanced. Finally, MMAPs were modified with 5-amino-benzimidazol (ABI) ligand and the adsorption capacity of IgG reached 153 mg/mL, higher than that of the normal magnetic particles (126 mg/mL). Moreover, adsorption behavior of MMAPs to IgG was little changed after twenty-five recycled use. Hence, MMAPs prepared herein showed great potential for bioseparation.
研究领域:
生物化学
·
材料科学
·
生物工程
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Biosensors-Basel
·
2025
·Vol. 15
·期 11
·747
·
IF 5.6
Qianyu Yang,Zhiwei Liu,Xinrui Xu,Zihao Zhao,Ze Fan,Bin Du,Jianjie Xu,Jiwei Xu,Jiang Wang,Bing Liu,Xihui Mu,Zhaoyang Tong
Over the past few decades, Fc fragment-conjugated proteins, such as Protein A, have been extensively utilized across a range of applications, including antibody purification, site-specific immobilization of antibodies, and the development of biosensing platforms. In this study, building upon our group prior research, we designed and screened an affinity DNA functional ligand (A-DNAFL) and experimentally validated its binding affinity (KD= 6.59 × 10−8) toward mouse IgG antibodies, whose binding performance was comparable to that of protein A. Systematic evaluations were performed to assess the binding efficiency under varying pH levels and ionic strength conditions. Optimal antibody immobilization was achieved in PBST-B buffer under physiological pH 7.2–7.4 and containing approximately 154 mM Na+and 4 mM K+. Two competitive binding assays confirmed that the A-DNAFL binds to the Fc fragment of murine IgG antibody. Furthermore, molecular docking simulations were employed to investigate the interaction mode, revealing key residues involved in binding as well as the contributions of hydrogen bonding and hydrophobic interactions to complex stabilization. Leveraging these insights, A-DNAFL was utilized as a tool for oriented immobilization of antibodies on the sensing interface, enabling the construction of an immunosensor for ricin detection. Following optimization of immobilization parameters, the biosensor exhibited a detection limit of 30.5 ng/mL with the linear regression equation is lg(Response) = 0.329 lg(Cricin) − 2.027 (N= 9,R= 0.938,p< 0.001)—representing a 64-fold improvement compared to conventional protein A-based methods. The system demonstrated robust resistance to nonspecific interference. Sensing interface reusability was also evaluated, showing only 8.55% signal reduction after two regeneration cycles, indicating that glycine effectively elutes bound antibodies while preserving sensor activity. In summary, the A-DNAFL presented in this study represents a novel antibody-directed immobilization material that serves as a promising alternative to protein A. It offers several advantages, including high modifiability, low production cost, and a relatively small molecular weight. These features collectively contribute to its broad application potential in biosensing, antibody purification, and other areas of life science research.
研究领域:
生物传感
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免疫学
·
分子识别
·
分析化学
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核酸工程
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Nanomaterials
·
2025
·Vol. 15
·期 8
·612
·
IF 4.3
Kang Yang,Fang Yang,Xiaoling Lu,Hao Li,Zeng Yang,Qi Yin,Lin Zhang,You Long,Chao Shen,Liya Chen,Bo Yao,Chenghong Huang
Simple and accurate analysis of cancer-related biomarkers is very important for disease screening and auxiliary diagnosis. This study proposed a facile immunoassay that used gold nanostar-labeled rabbit anti-AFP as a capture antibody and gold nanoparticle-conjugated goat anti-rabbit IgG as an enhance antibody for the construction of a detection strategy for AFP analysis. Investigations indicated that the 50 nm diameter GNS-labeled capture antibody can specifically catch AFPs by direct detection profile or by further signal amplification through AuNP-tagged enhance antibody combination. Results showed that the developed method holds 8.6 ng/mL sensitivity, 20.0–110.0 ng/mL detection range, acceptable precision and fine accuracy, as well as favorable specificity. Results of application to real serum determination by the proposed method are highly related to those of the ECLIA method (correlation coefficient is 0.931). The proposed method has simple-operation merit and is very suitable for clinical screening of large-scale serum samples of cancers.
研究领域:
医学诊断
·
纳米生物学
·
临床检验
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TALANTA
·
2015
·
IF 3.545
Di Zhang,Ying Sun,Qiong Wu,Pinyi Ma,Hua Zhang,Yuanpeng Wang,Daqian Song
A novel surface plasmon resonance (SPR) biosensor based on Ag nanocubes/chitosan composite was fabricated for mouse IgG detection. Ag nanocubes (AgNCs) were successfully synthesized with sulfide-mediated protocol. They were characterized with transmission electron microscopy (TEM) and UV–vis adsorption spectrum. AgNCs were etched by 3-Mercaptopropinic acid (MPA) and simply mixed with chitosan and glutaraldehyde . The Ag nanocubes/chitosan composite was deposited on Au film by the spin-coating. The electronic coupling between the AgNCs and the surface plasmon wave leads to the amplification of the SPR response, which results in the sensitivity enhancement of SPR biosensor. Moreover, antibody can be immobilized on Au film with aldehyde groups via Schiff alkali reaction. The traditional SPR biosensor based on MPA shows a response for mouse IgG in the concentration range of 2.50–40.00 μg mL −1 . The SPR biosensor based on AgNCs/chitosan composite shows a good response for mouse IgG in the concentration range of 0.60–40.00 μg mL −1 . The limit of quantification by the biosensor based on AgNCs/chitosan composite substrate is about 4 times lower than traditional biosensor, which has proved the SPR biosensor here proposed with more sensitivity and better performance than traditional SPR biosensor.
研究领域:
生物传感器
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纳米技术
·
分析化学
·
生物医学工程
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Nanoscale
·
2019
·Vol. 11
·期 12
·5526-5534
·
IF 6.97
Yu-Fang Ma,Li-Juan Wang,Ying-Lin Zhou,Xin-Xiang Zhang
The development of facilely synthetic materials for highly efficient enrichment of N-linked glycopeptides is essential in glycoproteome analysis. In this work, by utilizing the self-assembling of glutathione (GSH) on silver nanoparticles (Ag NPs), and the formation and dispersion of Ag NPs on a robust TpPa-1 substrate, a newly functionalized covalent organic framework (COF) called TpPa-1@Ag@GSH was synthesized via a simple two step post-synthetic modification. TpPa-1@Ag@GSH and intermediate products were confirmed and evaluated by nuclear magnetic resonance spectroscopy, Fourier transform infrared spectroscopy, X-ray diffraction, transmission electron microscopy, scanning electron microscopy-energy dispersive spectroscopy, Brunauer-Emmett-Teller and thermogravimetric analyses. Benefiting from the judicious selection of the substrate, the abundance of binding sites, relatively high affinity between GSH and N-linked glycopeptides, and the multivalent interactions between N-linked glycopeptides and unoccupied surfaces of Ag NPs, this porous material showed great performance in N-linked glycopeptide enrichment. By enriching N-linked glycopeptides in tryptic digests of human serum immunoglobulin G (human IgG) followed by mass spectrometry analysis, our method was proved to have good sensitivity (1 fmol), high selectivity (1 : 1500, human IgG to bovine serum albumin), high binding capacity (160 mg g−1, IgG/TpPa-1@Ag@GSH), ultra-fast capture ability (only 1 min incubation time), and good reusability (at least 5 times). It was also successfully applied to the enrichment of N-linked glycopeptides from complex biological samples. Our work improved the enrichment selectivity of COFs, reached the most rapid capture ability among off-column enrichment materials, and provided a very facile and easily popularized post-synthetic modification route for COFs in glycoproteome analysis.
研究领域:
分析化学
·
蛋白质组学
·
生物医学
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ANALYTICAL AND BIOANALYTICAL CHEMISTRY
·
2015
·Vol. 408
·期 4
·1015-1022
·
IF 3.436
Wang Shasha,Chen Zhaopeng,Choo Jaebum,Chen Lingxin
A naked-eye sensitive ELISA-like assay was developed based on gold-enhanced peroxidase-like activity of gold nanoparticles (AuNPs). Using human IgG (H-IgG) as an analytical model, goat anti-human IgG antibody (anti-IgG) adsorbed on microtiter plate and AuNPs-labeled anti-IgG acted as capture antibody and detection antibody, respectively. Because the surfaces of AuNPs were blocked by protein molecules, the peroxidase-like activity of AuNPs was almost inhibited, evaluated by the catalytic oxidation of peroxidase enzyme substrate 3,3′,5,5′-tetramethylbenzidine (TMB), which could produce a bright blue color in the presence of H 2 O 2 . Fortunately, the catalytic ability of AuNPs was dramatically increased by the deposition of gold due to the formation of a new gold shell on immunogold. Under optimal reaction conditions, the colorimetric immunoassay presented a good linear relationship in the range of 0.7–100 ng/mL and the limit of detection (LOD) of 0.3 ng/mL calculated by 3σ/S for UV-vis detection, and obtained LOD of 5 ng/mL for naked-eye detection. The obtained results were competitive with conventional sandwich ELISA with the LOD of 1.6 ng/mL. Furthermore, this developed colorimetric immunoassay was successfully applied to diluted human serum and fetal bovine serum samples, and predicted a broad prospect for the use of peroxidase-like activity involving nanomaterials in bioassay and diagnostics.
A naked-eye ELISA-like immunoassay for determination of human IgG is described based on gold enhanced peroxidase like activity of gold nanoparticles and it is a colorimetric typical sandwich-type immunoassay format with high sensitivity
研究领域:
分析化学
·
生物医学
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JOURNAL OF CHROMATOGRAPHY A
·
2023
·
IF 4.1
Guorong Ma,Ruiqiang Xu,Wang Pei,Yang Zhang,Rui Ma,Rang Yang,Zhihao Zhao,Yating Li,Chenjie Feng,Dongwu Jin,Wei Ma,Yumei Ma,Zhongren Ma
It is important to recycle the bovine blood discarded at slaughter and develop it into high value-added bovine serum products. Biomimetic affinity chromatography (BiAC) resins have been developed to specifically purify bovine serum immunoglobulin G (Bs-IgG). The BiAC strategy was used to screen the resins with the best purification effect on Bs-IgG. Four resins with specificity for Bs-IgG adsorption were selected from 90 BiAC resins. Finally, BiAC-A5–87 was selected and used to purify Bs-IgG based on the results of SDS-PAGE and BCA protein quantification analysis. The adsorption capacity and purity of BiAC-A5–87 were 32.79 ± 3.57 mg/mL and 85.9 ± 1.21 % for Bs-IgG, respectively. The total protein recovery rate of Bs-IgG purified by BiAC-A5–87 was 89.78±3.52 %. The resin of BiAC-A5–87 column was recycled in 40 breakthrough cycles, and its Bs-IgG adsorption efficiency decreased by less than 10 %. After soaking BiAC-A5–87 in 1.0 moL NaOH solution for 64 h, its adsorption capacity for Bs-IgG was almost the same as that before soaking. The development of waste bovine serum not only realizes the utilization of blood resources and produces high economic benefits but also reduces the pollution of the environment.
研究领域:
生物化学
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免疫学
·
蛋白质工程
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SENSORS AND ACTUATORS B-CHEMICAL
·
2019
·
IF 6.393
Ling-Xin Kong,Yan-Xin Zhang,Wei-Gang Zhang,Yun-Shan Zhang,Tie-Yi Yan,Peng-Cheng Geng,Biao Wang
Lab-on-tip technology opens a new door for optical fiber surface plasmon resonance (SPR) sensors. In this paper, we first demonstrate an ultra-compact reflective SPR microtip probe on the flat fiber end in experiment, which has a length of tens of microns and a diameter of several microns. Simulations and experiments show that this new fiber SPR microtip probe coated with a nano-goldfilm has excellent refractive index (RI) sensing characteristic and great potential for biochemical detection at single cell level. Based on the high RI sensitivity, a bio-probe assembled with Chitosan/Polysodium Styrene Sulfonate (Cs/PSS) nano-polyelectrolyte film and the sheep anti-human IgG-HRP is prepared to recognize human IgG specifically. Due to the high surface sensitivity, the SPR microtip probe exhibits an ultra-low detection limit for human IgG. Due to the high bulk sensitivity and high thermo-optical coefficient (TOC) of polydimethylsiloxane (PDMS), a reference temperature probe based on SPR microtip is proposed, which is used to compensate for the temperature cross-sensitivity of the bio-probe. This new SPR microtip probe can be used not only in optical fiber sensor, but also in Tip-Enhanced Near-Field Raman Microscopy and Scanning Near-Field Optical Microscope.
研究领域:
光纤传感器
·
表面等离子体共振(SPR)技术
·
生化传感
·
纳米技术
·
显微镜
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CHINESE JOURNAL OF ANALYTICAL CHEMISTRY
·
2010
·
IF 0.79
Guo-Feng XU,Hui LI,Cui ZHANG,Zi-Yin SUN,Wen-Ying ZHONG,Dan-Ke XU,Hong-Yuan CHEN
We synthesized and prepared a CdTe quantum dot (QD) probe modified with streptavidin (SA) and successfully established a novel visual detection method based on QD coupled with silver enhancement. Combined with the protein microarray analysis technology, this novel method did well in the detection of the reverse-phase protein microarray. To detect the analytical performance of this method, human IgG was used as a mode in this experiment. Initially, the human IgG was immobilized on aldehyde slides, then the optimized bio-goat anti-human IgG and CdTe-SA probe were added, and finally the silver enhancement coloration was held for 15 min; the results showed a good linear correlation between the signal gray level and the logarithm of human IgG immobilized on slides from 100 pg to 1.6 ng with a correlation coefficient ( R 2 ) of 0.997, while its lowest detectable limit was about 50 pg human IgG. This novel method could carry out the sensitive detection of microamounts of protein and shows the characteristics of a simple method that requires very few instruments, and the results obtained using this method can be easily visualized.
研究领域:
生物化学
·
分析化学
·
纳米技术
·
蛋白质组学
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TALANTA
·
2015
·
IF 3.545
Youming Shen,Youyu Zhang,Meiling Liu,Xiaoyin Liu,Huan Guo,Xiangyang Zhang,Chunxiang Zhang,Haitao Li,Shouzhuo Yao
In this work, a novel sulfur-containing compound with aldehyde groups was synthesized, which was used to immobilize antibodies through the covalent bonding of the aldehyde groups with amino groups, in which no additional chemical cross-linker is required. Human IgG was used as a model analyte to fabricate the electrochemical immunosensor. Using the proposed immunosensor, IgG was detected within the range from 0.01 to 25 ng mL −1 with a detection limit of 0.003 ng mL −1 obtained by 3 S/N. The simple electrochemical immunosensor had a good specificity, stability and reproducibility.
研究领域:
生物化学
·
免疫学
·
分析化学
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