JOURNAL OF CLINICAL LABORATORY ANALYSIS
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2024
·Vol. 38
·期 4
·e25011
·
IF 2.6
Ningning Wang,Qingqing Liu,Feihu Che,Qingyang Sun,Yue Wang,Chunli Yang,Yuzhu Dai,Jun Cheng
Background To establish a chemiluminescence method for detecting anti-E1 and anti-E2 antibodies in the serum of patients with hepatitis C virus (HCV) infection. Methods The microplate was coated with recombinant envelope proteins E1 and E2 by indirect method, respectively, and the kits for detecting anti-E1 and anti-E2 antibodies were prepared. The methodological indexes were evaluated. Results The methodological indexes of the kits were as follows: precision test (the variation coefficient of anti-E1 antibody 6.71%–8.95% for within run and 9.91%–12.16% for between run, the variation coefficient of anti-E2 antibody 6.06%–8.44% for within run and 10.77%–13.98% for between run, respectively). The blank limit and detection limit were 1.18 RLIR and 3.16 RLIR for the anti-E1 antibody, and 1.26 RLIR and 3.32 RLIR for the anti-E2 antibody, respectively. The correlation coefficients ( r ) of anti-E1 and anti-E2 were 0.9963 and 0.9828, the analysis and measurement ranges (AMR) were 1.66–41.28 RLIR and 1.55–19.46 RLIR, and the average recovery was 96.4% and 93.7%, respectively. The rheumatoid factor and other positive serum samples had no interference or cross-reaction to the test, and the kits were stable within 15 months. The positive rates of anti-E1 and anti-E2 antibodies in 45 patients with HCV infection were 35.6% (16/45) and 44.4% (20/45), respectively. Conclusions The kits for detecting anti-E1 and anti-E2 meet the requirements of methodology, and can be used in screening diagnosis, disease monitoring, prognosis evaluation, disease mechanism, and epidemiological studies of HCV infection. The HCV envelope proteins E1 and E2 have an immune response in HCV-infected patients.
CAS / MDL:
研究领域:
医学检验
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病毒学
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产品关联
DOI
PubMed
高影响因子
Nanoscale Research Letters
·
2014
·Vol. 9
·期 1
·1-13
·
IF 2.481
Li Chao,Ji Yang,Wang Can,Liang Shujing,Pan Fei,Zhang Chunlei,Chen Feng,Fu Hualin,Wang Kan,Cui Daxiang
Successful development of safe and highly effective nanoprobes for targeted imaging of in vivo early gastric cancer is a great challenge. Herein, we choose the CdSe/ZnS (core-shell) quantum dots (QDs) as prototypical materials, synthesized one kind of a new amphiphilic polymer including dentate-like alkyl chains and multiple carboxyl groups, and then used the prepared amphiphilic polymer to modify QDs. The resultant amphiphilic polymer engineered QDs (PQDs) were conjugated with BRCAA1 and Her2 monoclonal antibody, and prepared BRCAA1 antibody- and Her2 antibody-conjugated QDs were used for in vitro MGC803 cell labeling and in vivo targeted imaging of gastric cancer cells. Results showed that the PQDs exhibited good water solubility, strong photoluminescence (PL) intensity, and good biocompatibility. BRCAA1 antibody- and Her2 antibody-conjugated QD nanoprobes successfully realized targeted imaging of in vivo gastric cancer MGC803 cells. In conclusion, BRCAA1 antibody- and Her2 antibody-conjugated PQDs have great potential in applications such as single cell labeling and in vivo tracking, and targeted imaging and therapeutic effects' evaluation of in vivo early gastric cancer cells in the near future.
研究领域:
生物医学工程
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纳米医学
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肿瘤学
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DOI
PubMed
高影响因子
ANALYTICAL CHEMISTRY
·
2021
·
IF 6.986
Jia-Yang He,Xin Shang,Chun-Li Yang,Si-Yu Zuo,Ruo Yuan,Wen-Ju Xu
Exploring the ratiometric fluorescence biosensing of DNA-templated biemissive silver nanoclusters (AgNCs) is significant in bioanalysis, yet the design of a stimuli-responsive DNA device is a challenge. Herein, using the anti-digoxin antibody (anti-Dig) with two identical binding sites as a model, a tweezer-like DNA architecture is assembled to populate fluorescent green- and red-AgNCs (g-AgNCs and r-AgNCs), aiming to produce a ratio signal via specific recognition of anti-Dig with two haptens (DigH). To this end, four DNA probes are programmed, including a reporter strand (RS) dually ended with a g-/r-AgNC template sequence, an enhancer strand (ES) tethering two same G-rich tails (G18), a capture strand (CS) labeled with DigH at two ends, and a help strand (HS). Initially, both g-AgNCs and r-AgNCs wrapped in the intact RS are nonfluorescent, whereas the base pairing between RS, ES, CS, and HS resulted in the construction of DNA mechanical tweezers with two symmetric arms hinged by a rigid “fulcrum”, in which g-AgNCs are lighted up due to G18 proximity (“green-on”), and r-AgNCs away from G18 are still dark (“red-off”). When two DigHs in proximity recognize and bind anti-Dig, the conformation switch of these tweezers resultantly occurs, taking g-AgNCs away from G18 for “green-off” and bringing r-AgNCs close to G18 for “red-on”. As such, the ratiometric fluorescence of r-AgNCs versus g-AgNCs is generated in response to anti-Dig, achieving reliable quantization with a limit of detection at the picomolar level. Based on the fast stimulated switch of unique DNA tweezers, our ratiometric strategy of dual-emitting AgNCs would provide a new avenue for a variety of bioassays.
研究领域:
生物分析
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纳米生物学
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分子诊断
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DOI
PubMed
高影响因子
FOOD CHEMISTRY
·
2024
·
IF 8.5
Zhihong Xuan,Wenjie Shen,Hongmei Liu,Baoxia Ni,Ziye Lian,Li Li,Jinnan Chen,Baoyuan Guo,Songxue Wang,Jin Ye
This study presents a novel approach toward the one-pot green synthesis of ZIF-8/IgG composite, focusing on its precise orientation and protection of the anti-aflatoxins antibody. The antibody orientation is achieved through the specific binding of IgG to the Fc region of the antibody, while the antibody protection is accomplished by the structural change restriction of ZIF-8 framework to the antibody. Consequently, the antibody exhibits enhanced target capability and significantly improved tolerance to organic solvents. The ZIF-8/IgG/anti-AFT was employed for the purification and detection of AFTs by coupling with UPLC. Under optimized conditions, the recoveries of spiked AFTs in peanut oils are between 86.1% and 106.4%, with relative standard deviations (RSDs) ranging from 0.8% to 8.8%. The linearity range is 0.5–20.0 ng for AFB 1 and AFG 1 , 0.125–5.0 ng for AFB 2 and AFG 2 , the limit of detection is 0.1 ng for AFB 1 and AFG 1 , 0.03 ng for AFB 2 and AFG 2 .
研究领域:
生物化学
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食品科学
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环境分析
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产品关联
DOI
PubMed
高影响因子
FOOD CHEMISTRY
·
2024
·
IF 8.5
Jinkui Wang,Tong Bu,Qianqian Cui,Zizhe Li,Qing Xu,Zhixian Jiang,Yexuan Mao,Xianqing Huang,Lianjun Song,Dapeng Peng,Xiya Zhang
Simultaneous detection of carbofuran (CBF) and 3-hydroxy carbofuran (3-OH-CBF) in fruits and vegetables is important due to their high toxicity and widespread use in pest control. However, most lateral flow immunoassay (LFA) approaches only detection CBF. To overcome this limitation, two haptens, 6-((2,2-dimethyl-2,3-dihydrobenzofuran-7-yl)oxy)hexanoic acid and 6-((((2,2-dimethyl-2,3-dihydroben zofuran-7-yl)oxy)carbonyl)amino)hexanoic acid, named H1 and H2, were designed for broad-spectrum antibody detection of CBF and 3-OH-CBF. The highly specific monoclonal antibody (mAb) 3F4 based on hapten H1 only recognized CBF, while broad-spectrum mAbs 1D3, 6B7 and 6E8 based on hapten H2 recognized both compounds. The carbon bond to the hydroxyl group of 3-OH-CBF adopts a 28.55° angle relative to the same carbon bond of CBF, explaining the specificity of the H1-based mAb against CBF. After optimizing the antigen-antibody combination, a sensitive colloidal gold-LFA (CG-LFA) strip was constructed based on mAb 1D3 and H2-BSA for CBF and 3-OH-CBF detection following simple sample pretreatment. The established LFA had a limit of detection of 0.49–6.63 ng/mL, and adequate recoveries (69.3–105.7 %) for CBF and 3-OH-CBF in 26 fruit and vegetable samples.
研究领域:
环境科学
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食品安全
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免疫学
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DOI
PubMed
高影响因子
Biotechnology Journal
·
2024
·Vol. 19
·期 3
·2300688
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IF 3.2
Qing Li,Jianyu Zhu,Ting Yuan,Leina Dou,Xuezhi Yu,Jianzhong Shen,Huijuan Yang,Zhanhui Wang
Filamentous bacteriophage display technology has been employed in antibody discovery, drug screening, and protein–protein interaction study across various fields, including food safety, agricultural pollution, and environmental monitoring. Antifilamentous bacteriophage antibodies for identifying filamentous bacteriophage are playing a pivotal role in this technology. However, the existing antifilamentous bacteriophage antibodies lack sensitivity and specificity, and the antibodies preparation methods are cumbersome and hyposensitive. The major coat protein pVIII of filamentous bacteriophage has an advantage in quantification, which is benefit for detecting signal amplification but its full potential remains underutilized. In this study, the partial polypeptide CT21 of the major coat protein pVIII of filamentous bacteriophage was intercepted as the targeted immunogen or coating antigen to prepare antifilamentous bacteriophage antibodies. Six filamentous bacteriophage-specific monoclonal antibodies (mAbs) M5G8, M9A2, P6B5, P6D2, P8E4, and P10D4 were obtained. The limit of detections of the prepared six mAbs for detecting filamentous bacteriophage was 1.0 × 10 7 pfu mL −1 . These mAbs stayed stable under different pH, temperature, and exhibited high specificity in real application. This study not only provides a new idea for simplifying the preparation of antifilamentous bacteriophage antibodies which could apply in filamentous bacteriophage display, but it also presents a novel strategy for preparing antibodies against protein-specific epitopes with high sensitivity.
研究领域:
生物技术
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免疫学
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分子生物学
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DOI
PubMed
高影响因子
SENSORS AND ACTUATORS B-CHEMICAL
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2024
·
IF 8
Zizhe Li,Qianqian Cui,Qingyue Li,Changwei Luo,Mengtian Chen,Beibei Feng,Huan Li,Tong Bu,Yexuan Mao,Meng Dang,Xianqing Huang,Lianjun Song,Dapeng Peng,Xiya Zhang
To overcome limitations with respect to the low sensitivity of rapid immunoassay methods due to dilution of the extract in extraction solution, ultra-sensitive antibodies are needed. The linker-tethering site between the target and carrier protein , the length of the spacer arm , and the atomic charge of the linker-tethering site between the target and spacer arm in haptens design play a pivotal role in the preparation of ultra-sensitive antibodies. In this study, carbendazim (CBZ), a systemic broad-spectrum fungicidal pesticide, was used as model compound. 2-Aminobenzimidazole was chosen as the common skeletal structure and reacted with different groups, such as alkanes and urine groups, to form haptens H1 (previously reported), H2 (novel), and H3 (novel), respectively. Subsequently, eight monoclonal antibodies (mAbs) were generated using these three haptens , which exhibited notable variation in sensitivity. The half maximal inhibitory concentration (IC 50 ) value of mAb 4B11 based on hapten H3 against CBZ was 0.04 ng/mL, which was seven times better than that of mAb 5B10 (IC 50 value of 0.28 ng/mL) prepared using hapten H2 and 129 times better than that of mAb 6D5 (IC 50 value of 5.15 ng/mL) prepared using hapten H1. Finally, the optimal antigen–antibody combination was employed to establish a sensitive colloidal gold lateral flow immunoassay (CG-LFA) for CBZ detection in vegetables and fruits with convenient sample pretreatment . The developed CG-LFA shows promising practical utility and prospects for application due to its low limit of detection (0.10–0.18 ng/mL) and high recovery ratios (76.0%–96.1%) in six different samples.
研究领域:
农业科学
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免疫学
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分析化学
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DOI
高影响因子
JOURNAL OF CHROMATOGRAPHY B-ANALYTICAL TECHNOLOGIES IN THE BIOMEDICAL AND LIFE SCIENCES
·
2016
·
IF 2.687
Wenhua Li,Bin Yang,Dongmei Zhou,Jun Xu,Zhi Ke,Wen-Chen Suen
Liquid chromatography mass spectrometry (LC–MS) is the most commonly used technique for the characterization of antibody variants. MAb-X and mAb-Y are two approved IgG1 subtype monoclonal antibody drugs recombinantly produced in Chinese hamster ovary (CHO) cells. We report here that two unexpected and rare antibody variants have been discovered during cell culture process development of biosimilars for these two approved drugs through intact mass analysis. We then used comprehensive mass spectrometry-based comparative analysis including reduced light, heavy chains, and domain-specific mass as well as peptide mapping analysis to fully characterize the observed antibody variants. The “middle-up” mass comparative analysis demonstrated that the antibody variant from mAb-X biosimilar candidate was caused by mass variation of antibody crystalline fragment (Fc), whereas a different variant with mass variation in antibody antigen-binding fragment (Fab) from mAb-Y biosimilar candidate was identified. Endoproteinase Lys-C digested peptide mapping and tandem mass spectrometry analysis further revealed that a leucine to glutamine change in N-terminal 402 site of heavy chain was responsible for the generation of mAb-X antibody variant. Lys-C and trypsin coupled non-reduced and reduced peptide mapping comparative analysis showed that the formation of the light-heavy interchain trisulfide bond resulted in the mAb-Y antibody variant. These two cases confirmed that mass spectrometry-based comparative analysis plays a critical role for the characterization of monoclonal antibody variants, and biosimilar developers should start with a comprehensive structural assessment and comparative analysis to decrease the risk of the process development for biosimilars.
研究领域:
生物技术
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药物研发
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蛋白质组学
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产品关联
DOI
PubMed
高影响因子
ANALYTICAL CHEMISTRY
·
2021
·
IF 6.986
Hailing Zheng,Hailiang Yang,Yang Zhou,Tianxiao Li,Qinglin Ma,Bing Wang,Qin Fang,Haixiang Chen
The extraction and identification of silk residues in tombs is of great significance for studying the distribution and spread of early silk. However, the complex organic matter in the tomb hinders the accurate identification of silk. In this study, a double-antibody sandwich enzyme-linked immunosorbent assay (ELISA) based on immunomagnetic beads (IMBs) was developed for the rapid enrichment and detection of silk residues. The double-antibody sandwich ELISA method established by pairing the IMBs prepared by the silk fibroin monoclonal antibody SF-3 and the silk fibroin monoclonal-labeled antibody bio-SF-1 had the highest detection sensitivity, with a linear detection range of 10 to 104 ng mL–1 and a detection limit of 5.12 ng mL–1. This method was excellent in the extraction and analysis of silk residues from archaeological imprints and soil samples and successfully identified silk residues in samples at the final stage of silk degradation (physical invisible silk). The proteomics analysis results demonstrated the feasibility and practicability of this method.
研究领域:
考古学
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生物化学
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材料科学
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产品关联
DOI
PubMed
高影响因子
Nature Communications
·
2021
·Vol. 12
·期 1
·1-14
·
IF 14.919
Jiang Cheng-Tao,Chen Kai-Ge,Liu An,Huang Hua,Fan Ya-Nan,Zhao Dong-Kun,Ye Qian-Ni,Zhang Hou-Bing,Xu Cong-Fei,Shen Song,Xiong Meng-Hua,Du Jin-Zhi,Yang Xian-Zhu,Wang Jun
Modulating effector immune cells via monoclonal antibodies (mAbs) and facilitating the co-engagement of T cells and tumor cells via chimeric antigen receptor- T cells or bispecific T cell-engaging antibodies are two typical cancer immunotherapy approaches. We speculated that immobilizing two types of mAbs against effector cells and tumor cells on a single nanoparticle could integrate the functions of these two approaches, as the engineered formulation (immunomodulating nano-adaptor, imNA) could potentially associate with both cells and bridge them together like an ‘adaptor’ while maintaining the immunomodulatory properties of the parental mAbs. However, existing mAbs-immobilization strategies mainly rely on a chemical reaction, a process that is rough and difficult to control. Here, we build up a versatile antibody immobilization platform by conjugating anti-IgG (Fc specific) antibody (αFc) onto the nanoparticle surface (αFc-NP), and confirm that αFc-NP could conveniently and efficiently immobilize two types of mAbs through Fc-specific noncovalent interactions to form imNAs. Finally, we validate the superiority of imNAs over the mixture of parental mAbs in T cell-, natural killer cell- and macrophage-mediated antitumor immune responses in multiple murine tumor models.
研究领域:
癌症免疫疗法
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纳米技术
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免疫学
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产品关联
DOI
PubMed
高影响因子
ANALYTICA CHIMICA ACTA
·
2022
·
IF 6.911
Yarong Ren,Juan Wei,Ye Wang,Peng Wang,Yanwei Ji,Baoyuan Liu,Jianlong Wang,Gualberto González-Sapienza,Yanru Wang
Salmonella are major pathogens that cause foodborne diseases. In this work, a broad-spectrum Salmonella nanobody-01 (Nb-01) was isolated and applied in the development of a streptavidin-bridged sandwich ELISA (SAB-ELISA) for simultaneously identifying five Salmonella serovars , including Salmonella Enteritidis ( S. Enteritidis), Salmonella Typhimurium ( S. Typhimurium), Salmonella London (S. London), Salmonella Paratyphi B (S. Paratyphi B) and Salmonella Hadar (S. Hadar). In this work, streptavidin (SA) was utilized as a scaffold to directionally immobilize biotinylated nanobody (BiNb) and Salmonella was detected by phage-displayed nanobodies. The SAB-ELISA can be accomplished within 180 min with a limit of detection (LOD) of 6.31 × 10 3 colony forming units (CFU) mL −1 , 9.15 × 10 3 CFU mL −1 , 4.23 × 10 3 CFU mL −1 , 7.31 × 10 3 CFU mL −1 and 7.25 × 10 3 CFU mL −1 towards S. Typhimurium, S. Enteritidis, S. London, S. Paratyphi B and S. Hadar, respectively. In comparison of sandwich ELISA by passive immobilization of Nb-01 on polystyrene plate, the sensitivity was increased by around 6-fold, which confirmed the enhanced immobilization efficacy of SAB-ELISA. Furthermore, the feasibility of the assay for S. Typhimurium determination in actual samples was evaluated, showing excellent recovery, inter-day and intra-day precision.
研究领域:
微生物学
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免疫学
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食品科学
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分子生物学
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产品关联
DOI
PubMed
高影响因子
PLoS Neglected Tropical Diseases
·
2021
·Vol. 15
·期 3
·e0008403
·
IF 4.411
Yuan Zhang,Yanqiu Wei,Yunlong Li,Xuan Wang,Yang Liu,Deyu Tian,Xiaojuan Jia,Rui Gong,Wenjun Liu,Limin Yang
Ebola virus (EBOV) is one of the most virulent pathogens that causes hemorrhagic fever and displays high mortality rates and low prognosis rates in both humans and nonhuman primates. The post-exposure antibody therapies to prevent EBOV infection are considered effective as of yet. However, owing to the poor thermal stability of mammalian antibodies, their application in the tropics has remained limited. Therefore, a thermostable therapeutic antibody against EBOV was developed modelled on the poultry(chicken) immunoglobulin Y (IgY). The IgY antibodies retaining their neutralising activity at 25°C for one year, displayed excellent thermal stability, opposed to conventional polyclonal antibodies (pAbs) or monoclonal antibodies (mAbs). Laying hens were immunised with a variety of EBOV vaccine candidates and it was confirmed that VSVΔG/EBOVGP encoding the EBOV glycoprotein could induce high titer neutralising antibodies against EBOV. The therapeutic efficacy of immune IgY antibodies in vivo was evaluated in the newborn Balb/c mice who have been challenged with the VSVΔG/EBOVGP model. Mice that have been challenged with a lethal dose of the pseudovirus were treated 2 or 24 h post-infection with different doses of anti-EBOV IgY. The group receiving a high dose of 10 6 NAU/kg (neutralising antibody units/kilogram) showed complete protection with no symptoms of a disease, while the low-dose group was only partially protected. Conversely, all mice receiving naive IgY died within 10 days. In conclusion, the anti-EBOV IgY exhibits excellent thermostability and protective efficacy. Anti-EBOV IgY shows a lot of promise in entering the realm of efficient Ebola virus treatment regimens.
Despite the amount of efficient Ebola virus therapeutic antibodies reported in recent years, their application in tropical endemic areas has remained limited due to the low thermal stability of mammalian antibodies. A highly thermostable therapeutic polyclonal antibody against EBOV was developed based on chicken immunoglobulin Y (IgY). The EBOV specific IgY antibodies displayed excellent thermal stability, retaining their neutralising activity at 25°C for one year. The newborn mice receiving the passive transfer of IgY achieved complete protection against a lethal dose of virus challenge proving that the anti-EBOV IgY provides a promising recourse to solve some of the current clinical application hindrances of Ebola antibody-based treatments in Africa due to thermal stability.
研究领域:
病毒学
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免疫学
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治疗学
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传染病
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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）