TALANTA
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2017
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IF 4.162
Xiaoxiao Su,Mengyao Wang,Yue Wu,Yong He,Zhifeng Fu
A novel dual-site recognition protocol was developed for chemiluminescent (CL) detection of Streptococcus mutans ( S. mutans ) based on a designed antibiotic-affinity strategy. Teicoplanin , a broad-spectrum antibiotic against Gram-positive bacteria, was adopted to functionalize magnetic particles and recognize S. mutans utilizing the strong affinity between this agent and D-Alanyl-D-Alanine peptide moieties in the bacterial cell wall. To achieve ideal specificity for S. mutans detection, rat immunoglobulin G2a (rat IgG2a) tagged with horseradish peroxidase (HRP) was used as the second recognition agent and signal tracer since Fab region of rat IgG2a could bind with streptococcal protein G highly expressed in the cell wall of S. mutans . Thus HRP-tagged sandwich complex of teicoplanin/ S. mutans /rat IgG2a was formed on the magnetic particles, followed by a CL quantification of S. mutans based on a HRP-catalyzed luminol-H 2 O 2 -p -iodophenol CL reaction. This dual-site recognition protocol showed a linear range of 1.0 × 10 2 –1.0 × 10 6 CFU mL −1 and a detection limit of 33 CFU mL −1 for S. mutans detection. The whole detection process could be completed within 70 min. The recovery tests for food, environmental, pharmaceutical and biological samples showed acceptable recovery values between 83.0% and 110.0%, demonstrating its application potential for detection of bacteria in various sample matrixes.
研究领域:
微生物学
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分析化学
✓ 已验证
产品关联
DOI
PubMed
高影响因子
SENSORS AND ACTUATORS B-CHEMICAL
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2017
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IF 5.401
Shijia Yang,Mengyao Wang,Lin Wang,Zhifeng Fu
A sandwich protocol was developed for chemiluminescent detection of Streptococcus mutans ( S. mutans ) utilizing dual-site recognition of the bacteria by rat IgG2a and vancomycin. Magnetic particles (MPs) functionalized with rat IgG2a were adopted to capture S. mutans based on the binding between Fc region of rat IgG2a and protein G in the cell wall of the bacteria. Alkaline phosphatase labeled vancomycin was applied as the signal tracer since D-Ala- d -Ala peptide moieties in the cell wall of Gram-positive bacteria could be recognized by this antibiotic agent. The protocol showed an ideal specificity for S. mutans detection because the recognition agents bound with the bacteria at two distinct sites. Furthermore, the utilization of MPs enrichment greatly improved the sensitivity. S. mutans could be detected within a linear range of 2.4 × 10 1 − 2.4 × 10 5 CFU mL −1 , with a very low detection limit of 2.3 × 10 1 CFU mL −1 . Some common Gram-positive bacteria and Gram-negative bacteria showed negligible interference. This proposed method for S. mutans detection possessed some attractive characteristics such as facile manipulation, short detection time, low cost, ultra-high sensitivity and ideal specificity. It was used to detect S. mutans in human saliva with acceptable recoveries ranging from 70.8% to 108.3%.
研究领域:
微生物学
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生物分析化学
✓ 已验证
产品关联
DOI
高影响因子
Jove-Journal of Visualized Experiments
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2023
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IF 1.2
Xiaoqiang Zeng,Hongwu Sun,Yan Ye,Xing Luo,Dingyi Cai,Yun Yang,Ting Chen,Cun Sun,Shaotong Zhang,Hao Zeng
Nanoemulsion adjuvant vaccines have attracted extensive attention because of their small particle size, high thermal stability, and ability to induce validly immune responses. However, establishing a series of comprehensive protocols to evaluate the immune response of a novel nanoemulsion adjuvant vaccine is vital. Therefore, this article features a rigorous procedure to determine the physicochemical characteristics of a vaccine (by transmission electron microscopy [TEM], atomic force microscopy [AFM], and dynamic light scattering [DLS]), the stability of the vaccine antigen and system (by a high-speed centrifuge test, a thermodynamic stability test, SDS-PAGE, and western blot), and the specific immune response (IgG1, IgG2a, and IgG2b). Using this approach, researchers can evaluate accurately the protective effect of a novel nanoemulsion adjuvant vaccine in a lethal MRSA252 mouse model. With these protocols, the most promising nanoemulsion vaccine adjuvant in terms of effective adjuvant potential can be identified. In addition, the methods can help optimize novel vaccines for future development.
研究领域:
疫苗学
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免疫学
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微生物学
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药学
✓ 已验证
产品关联
DOI
PubMed
高影响因子
INTERNATIONAL JOURNAL OF BIOLOGICAL MACROMOLECULES
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2025
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IF 8.5
Changcai Teng,Wenjing Ma,Jinfeng Liu,Juan Hou,Yalong Zhang,Xiongyan Meng,Yannan Xue,Zhen Wang,Jiajia Wang,Dexiang Chen,Qiang Sui,Qi Gao,Xia Li,Tiehai Li,Chengli Zong
Aberrantly glycosylated Mucin 1 (MUC1) is frequently over-expressed in epithelial cancers, making it an attractive target for cancer vaccines. Over the past two decades, multiple MUC1-based vaccines have been investigated clinically, yet they have generally shown limited efficacy due to challenges such as low immunogenicity, difficulty in overcoming immune tolerance, and potential issues related to glycosylation effects and antigen presentations. To advance the understanding of MUC1 vaccines, we report an efficient chemo-enzymatic approach for the preparation of four MUC1 antigen variants with different glycoforms through liquid-phase glycopeptide synthesis. These antigen were conjugated with CRM197 to generate various glycopeptide-protein conjugate vaccines, and their immunogenicity was evaluated based on total and subtype antibody titers, binding affinity, complement-dependent cytotoxicity (CDC) activity, and antibody-dependent cellular phagocytosis (ADCP). The combination of MPL and QS21 adjuvants with STn-MUC1-CRM197 conjugate induced a potent Th1-biased immune response, evidenced by elevating IgG2a titers and stronger antibody binding to MCF-7 cells. This formulation demonstrated superior CDC activity, ADCP activity and binding affinity to human breast cancer tissues in immuno-histochemical assays. The synergistic effect of specific adjuvants and glycosylated MUC1 conjugates offers a strategic avenue for MUC1 cancer vaccine development.
研究领域:
癌症研究
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免疫学
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化学
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药学
✓ 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）