Journal of Immunotoxicology
·
2022
·
IF 3.439
Wenhua Zhong,Penghuan Chang,Lianfang Gan,Lifan Zhong,Zhaoxin Yang
Most current methods to assess T-cell-dependent antibody responses (TDAR) are semi-quantitative and based on measures of antibody titer generated against a standard antigen like keyhole limpet hemocyanin (KLH). The precision, sensitivity, and convenience of TDAR assays might be improved by applying rapid, sensitive, specific cytometric bead assays (CBA). In the study here, KLH antigen was covalently coupled onto the surface of cytometric beads using immune microsphere technology, and IgM antibody capture spheres were prepared for use in pretreatment processing of samples. The working parameters associated with this novel TDAR-CBA system were optimized in orthogonal experiments. The optimal concentration of the KLH coating solution in this system was 160 μg/ml, that of the anti-KLH IgG capture spheres 6.0 × 105/ml, and the optimal dilution of fluorescein isothiocyanate (FITC)-conjugated Affini-Pure Goat Anti-Mouse IgG (H + L) was 60 μg/ml. Repeated tests indicated that this approach yielded good linearity (r2 = 0.9937) method, with a within-run precision of 3.1–4.9%, and a between-run precision of 4.4–4.9%. This new approach had a limit of detection of 113.43 ng/ml (linear range = 390.63–50 000), and an interference rate of just 0.04–3.51%. Based on these findings, it seems that a new mouse TDAR assay based on CBA can be developed that would appear to be more sensitive, accurate, and precise than the current TDAR assay approaches based on traditional ELISA.
研究领域:
免疫学
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分子生物学
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分析化学
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PubMed
高影响因子
Frontiers in Microbiology
·
2018
·
IF 4.019
Yu Guanliu,Yu Xianglong,Yang Guoping,Tang Yi,Diao Youxiang
Duck Tembusu virus (DTMUV) is an emerging pathogenic flavivirus that has resulted in large economic losses to the duck-rearing industry in China since 2010. Therefore, an effective diagnostic approach to monitor the spread of DTMUV is necessary. Here, a novel diagnostic immunochromatographic strip (ICS) assay was developed to detect DTMUV. The assay was carried out using colloidal gold coated with purified monoclonal antibody A12D3 against envelope E protein. Purified polyclonal C12D1 antibodies from BALB/c mice against the envelope E protein were used as the capture antibody. Goat anti-mouse IgG was used to detect DTMUV, which was also assembled on the ICS. Results showed that the ICS could specifically detect DTMUV within 10 min. It also could be stored 25 and 4°C for 4 and 6 months, respectively. The sensitivity of the ICS indicated that the dilution multiples of positive allantoic fluid of DTMUV (LD50: 104.33/0.2 ml) was up to 200. Its specificity and sensibility showed no significant change under the above storage situations. Fifty clinical samples were simultaneously detected by ICS and reverse-transcription polymerase chain reaction with a 93.9% coincidence rate between them. It proved that the ICS in the present study was highly specific, sensitive, repeatable, and more convenient to rapidly detect DTMUV in clinical samples.
研究领域:
兽医学
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病毒学
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诊断技术
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PubMed
高影响因子
Journal of Immunotoxicology
·
2019
·
IF 1.734
Penghuan Chang,Ling Huang,Mianqing Huang,Shuhong Tian,Zhaoxin Yang
Although T-cell-dependent antibody response (TDAR) assays with keyhole limpet hemocyanin (KLH) as specific antigen have many advantages, most experiments produce qualitative results based on antibody titers. It was hypothesized that if experimental conditions (like antigen coating concentration, serum dilution, and detecting [here, horseradish peroxidase-goat anti-mouse IgG] antibody dilution) could be optimized, the resulting measured value (here, optical density) could be used to directly analyze and evaluate the experimental results. This means specifically that the assay OD values could be used for approximate quantitative statistical analysis; it does not require a further conversion of the data into qualitative forms or require obtaining further titer data from additional experiments. As such, the use of this “improved” assay would: greatly reduce the complexity of experimental operations; improve overall sensitivity and practicality of traditional TDAR assays; and, allow for direct assessing of any immunosuppression caused by a test drug in a host. Here, KLH-immunized serum was obtained from BALB/c mice, and the means to detect serum anti-KLH antibodies by an indirect ELISA were optimized. The results indicated that in this system, the optimal KLH coating concentration was 80 μg/ml, the optimal dilution range of the serum (at immunization dose of 5 mg KLH/kg) was 1:200–1:800, and the optimal dilution of HRP-goat anti-mouse IgG antibody was 1:16,000. Methodology verification was performed and a regression model of y = 144.16x + 0.9815 (R2 = 0.9571, indicating very good linearity) was obtained. Intragroup precision was 7.51–9.40%; the intergroup coefficient of variation was 9.83–14.22%. The lower limit of detection was 0.1385. The present results showed this indirect ELISA exhibited very good linearity, accuracy, and precision.
研究领域:
免疫学
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生物化学
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药理学
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PubMed
高影响因子
SENSORS AND ACTUATORS B-CHEMICAL
·
2023
·
IF 8.4
Cuixia Guo,Ziling Ye,Yonghong He,Zeyu Huang,Hongjian Zhang,Feng Huang
In response to the urgent requirement for high-throughput multiplexed detection, a novel dual-encoded suspension array (SA) based on thickness and digital fluorescent peak is proposed, along with the corresponding optical system. The SAs are prepared by thickness encoded micro-glass chips (MGCs) with electrostatically adsorbed quantum dots (QDs). Using the thickness characteristics of microcarrier itself and digital fluorescence peak (DFP) as the encoded signals ensure the high encoding stability with simple fabrication process at a low cost. A spectral-domain phase and fluorescence spectroscopy microscopy (SDPFSM) system is designed for the first time to decode the thickness information and DFP with ultrahigh accuracy. The high selectivity and sensitivity of the dual-encoded SAs are verified by the multiplexed immunoassays . The quantitative detection of anti-human IgG, anti-mouse IgG, and anti-rabbit IgG was realized with the limits of detection (LOD) of 0.19, 0.35 and 0.31 pM/mL and the sensitivity of 713, 470 and 456 a.u./(pM/mL), respectively. The coding number increases by a factor of 458 compared with the traditional fluorescence coding method due to the additional use of the thickness information of the MGCs itself as a coding channel. The aforementioned properties demonstrate that the dual-encoded SA and the corresponding decoding platform are promising and robust in multiplexed detection.
研究领域:
生物传感器
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分析化学
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生物医学工程
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光学工程
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高影响因子
MICROCHIMICA ACTA
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2014
·Vol. 182
·期 3
·855-862
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IF 3.719
Yan Fufeng,Zhang Yuanchang,Zhang Shuai,Zhao Jihong,Liu Shunli,He Linghao,Feng Xiaozhong,Zhang Hongzhong,Zhang Zhihong
We report on an efficient method for the production carboxy-modified graphene (carboxy-GR), and its application to a highly sensitive immunoassay for the livestock feed additive clenbuterol. Octadecanoic acid is shown to self-assemble on carboxy-GR via intermolecular interaction. Goat anti-mouse IgG was conjugated to 1-octadecanethiol-modified carboxy-GR and then deposited by self-assembly on a C18-modified gold surface. The resulting sensor surface was used to quantify clenbuterol by both surface plasmon resonance and electrochemical impedance spectroscopy. The assay works in the 0.01 ng∙mL −1 to 10 ng∙mL −1 concentration range and the lower detection limit is 6.57 pg∙mL −1 . It is concluded that this fairly simple method can be extended to immunoassays for other food additives.
Principle of sensor system and immunoassay. (i) Binding of ACG–IgG with the alkyl chain of 1-octadecanethiol through intermolecular force onto the gold substrate; (ii) absorbance of Ab CLB onto the surface of ACG-IgG; and (iii) detection of CLB using an immunoassay based on Ab CLB –ACG–IgG.
研究领域:
材料科学
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分析化学
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食品安全
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生物化学
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纳米技术
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DOI
高影响因子
FOOD AND AGRICULTURAL IMMUNOLOGY
·
2018
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IF 2.568
Wenliang Ge,Steven Suryoprabowo,Xiaoling Wu,Qiankun Zheng,Hua Kuang
The misuse of β-agonists constitutes a potential risk to public health and β-agonists has been forbidden in many countries. In this study, we describe an immunochromatographic test strip method for the rapid, sensitive, and inexpensive detection of mabuterol (MAB) and mapenterol (MAP) by cross-reactivity in pig urine. For the immunochromatographic test, the coating antigen which was obtained by conjugating the salbutamol to bovine serum albumin and a goat anti-mouse IgG antibody were drawn onto a nitrocellulose membrane as the control line and test line. The test was developed with a sensitive monoclonal antibody specific for MAB, which was generated by immunizing BALB/c mice with a well-characterized MAB- keyhole limpet haemocyanin conjugate, produced in our laboratory. Under optimized conditions, the cut-off limits of the semi-quantitative test strip for MBA and MPA were as low as 10 ng/mL in both 0.01 M PBS (pH 7.4) and pig urine. All the results were obtained within 5 min. Our data indicate that the immunochromatographic assay allows provide sensitive, rapid, and specific on-site screening for MAB and MAP.
研究领域:
兽医学
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分析化学
·
公共卫生
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高影响因子
MICROCHIMICA ACTA
·
2023
·Vol. 190
·期 3
·1-8
·
IF 5.7
Yu Yao,Li Yansong,Xu Ning,Zha Yonghong,Bai Xue,Liu Mingyuan,Zhou Yu,Liu Xiaolei
Fumonisin B1 (FB1) is one of the important mycotoxins posing health risks in the area of food safety. A sensitive fluorescence ratio immunoassay has been established for FB1 based on the growth of monodispersed 2-D MnO 2 nanosheet on an individual gold nanoparticle (AuNP@MnO 2 ). FB1 competed with the coated FB1-BSA to bind the FB1 monoclonal antibody. After a washing step, alkaline phosphatase-labeled goat anti-mouse IgG (ALP-IgG) with high catalytic activity was combined with FB1 monoclonal antibody. ALP reacts with ascorbic acid 2-phosphate (AAP) to produce ascorbic acid (AA), which decomposes AuNP@MnO 2 to dehydroascorbic acid (DHAA). O-Phenylenediamine dihydrochloride (OPD) is oxidized to yellow-fluorescent substrate of 2,3-diaminophenazine (DAP) (excitation, 423 nm; emission, 570 nm) by AuNP@MnO 2 . Meanwhile, OPD can also be reduced to blue fluorescent substrate of OPDred (excitation, 350 nm; emission, 430 nm) by DHAA. The content of FB1 can be determined by fluorescence ratio of blue/yellow. The limit of detection (LOD) of the fluorescence ratio immunoassay for FB1 was 0.06 ng mL −1 , and the linear range was from 0.25 to 60.00 ng mL −1 . The effectiveness of the assay was verified in real maize samples, and satisfactory recoveries were attained. The correlation coefficient of these results between the fluorescence ratio immunoassay and commercial ELISA kit was 0.9999. This method provides a sensitive and selective tool for the detection of FB1 in maize samples. Graphical Abstract
研究领域:
食品安全
·
分析化学
·
纳米技术
·
毒理学
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DOI
PubMed
高影响因子
Journal of Materials Chemistry B
·
2024
·
IF 6.1
Zhiheng Zhang,Hongchen Chen,Weize Diao,Chuangxin Zhou,Guozhen Liu
Eutectic gallium indium (EGaIn) nanoparticles can be modified with aniline derivatives to introduce versatile functional groups toward bioapplications beyond materials science. The p-aminobenzoic acid (PABA) modified EGaIn (EGaIn-PABA) demonstrated high wettability with the presence of carboxyl groups, but the priority of carboxyl and amino groups binding on EGaIn surface remains unclear. To study the binding nature of PABA to EGaIn, the anti-mouse IgG antibody labeled with fluorescein isothiocyanate (FITC) (IgG-FITC) was covalently modified to EGaIn-PABA to verify the presence of terminal carboxyl groups on EGaIn surface. The binding of gold nanoparticles (AuNPs) to EGaIn-PABA nanoparticles suggested the presence of terminal amino groups on EGaIn surface. Then, taking advantage of the reductive nature of amino groups, the Almar blue fluorescence experiment was designed to determine the co-existence of carboxyl and amino groups on the EGaIn-PABA surface with the approximate ratio of 3:7, suggesting carboxyl groups had a higher probability of binding with the EGaIn surface than that of amino groups. Then, an aptasensor was fabricated on the EGaIn-PABA surface with AuNPs for electrochemical detection of interleukin-6 with a sensitivity of 1 pg/mL.
研究领域:
材料科学
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生物传感器
·
纳米技术
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分析化学
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生物医学工程
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DOI
PubMed
高影响因子
TALANTA
·
2023
·
IF 6.1
Gaozhen Li,Jiashuai Sun,Jiahuan Li,Yaoli Zhang,Jingcheng Huang,Fengling Yue,Haowei Dong,Falan Li,Huihui Xu,Yanyin Guo,Yemin Guo,Xia Sun
In order to achieve a highly sensitive detection of procymidone in vegetables, three paper-based biosensors based on a core biological immune scaffold (CBIS) were developed, which were time-resolved fluorescence immunochromatography strips with Europium (III) oxide (Eu-TRFICS). Goat anti-mouse IgG and europium oxide time-resolved fluorescent microspheres formed secondary fluorescent probes. CBIS was formed by secondary fluorescent probes and procymidone monoclonal antibody (PCM-Ab). The first type of Eu-TRFICS (Eu-TRFICS-(1)) fixed secondary fluorescent probes on a conjugate pad, and PCM-Ab was mixed with a sample solution. The second type of Eu-TRFICS (Eu-TRFICS-(2)) fixed CBIS on the conjugate pad. The third type of Eu-TRFICS (Eu-TRFICS-(3)) was directly mixed CBIS with the sample solution. They solved the problems of steric hindrance of antibody labeling, insufficient exposure of antigen recognition region and easy loss of activity in traditional methods. They realized multi-dimensional labeling and directional coupling. They replaced the loss of antibody activity. And the three types of Eu-TRFICS were compared, among which Eu-TRFICS-(1) was the best detection choice. Antibody usage was reduced by 25% and sensitivity was increased by 3 times. Its detection range was 1–800 ng/mL, the limit of detection (LOD) was 0.12 ng/mL with the visible LOD (vLOD) of 5 ng/mL.
研究领域:
分析化学
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生物传感
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食品安全
·
环境监测
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产品关联
DOI
PubMed
高影响因子
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