Frontiers in Bioengineering and Biotechnology
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2025
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IF 4.8
Guangwei Zhao,Shengshuo Zhang,Yujie Zou,Nan Jia,Liyuan Zheng,Jinhua Dong
Introduction: Testosterone is a steroid hormone, which plays a pivotal role in regulating metabolism and protein synthesis in the body. The detection of testosterone is of paramount importance for diagnostic purposes in clinical settings, as well as for monitoring athletes’ physiological parameters and ensuring the integrity of sports competitions.Methods: Herein, we present a phage display-derived biosensing platform through genetic engineering of the TS77 antibody variable domains. The variable region genes of the heavy and light chains from TS77 antibody were cloned into the pDong1 plasmid and displayed on the phage surfaces through phage display technology. Subsequently, a novel non-competitive open-sandwich ELISA (OS-ELISA) was developed for testosterone detection, leveraging the antigen-induced interaction changes in antibody variable regions.Results: OS-ELISA based on anti L-chain antibody achieved a limit of detection (LOD) of 2.71 nM and a half-maximal effective concentration (EC50) of 0.22 μM for testosterone detection. Furthermore, the enhanced OS-ELISA platform incorporating purified maltose binding protein fused with VL (MBP-VL) and VH phage demonstrated a LOD of 1.07 pM and a wide working range from 1 pM to 10 mM.Discussion: The OS-ELISA developed in this study exhibits high sensitivity and a broad dynamic range for testosterone quantification, showing significant potential for clinical diagnostics and athlete monitoring applications.
研究领域:
生物化学
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免疫学
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临床诊断
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DOI
PubMed
高影响因子
MICROCHEMICAL JOURNAL
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2024
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IF 4.9
Yufan Zhang,Li Luo,Siyue Tian,Bingjian Zhang,Qiang Li
Archaeobotany plays a vital role in understanding the origins and development of agriculture. Given the different primary crops in northern and southern China, developing a method for distinguishing between millet and rice residues is crucial. Enzyme-linked immunosorbent assay (ELISA) has unique advantages in detecting animal organic residues, but its effectiveness in detecting plant residues remains uncertain. Therefore, this study devised an optimized ELISA method for the rapid, high-throughput identification of millet and rice residues in archaeological samples. The method exhibited satisfactory sensitivity and specificity, effectively detecting plant organic residues in both categories of archaeological samples. The method’s applicability and accuracy were cross-validated through documentary records, iodine–potassium iodide testing, and phytolith analysis. This study also represents a preliminary application of ELISA in archaeobotany. With further research, this technology has the potential to become a valuable tool in plant archaeology.
研究领域:
植物考古学
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生物化学
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DOI
高影响因子
MICROCHIMICA ACTA
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2025
·Vol. 192
·期 9
·1-13
·
IF 5.3
Kang Jia,Qi Meng,Wang Yasen,Li Linchun,Zhou Xinda,An Xiaoshuo,Hammock Bruce D.,Zhang Jinlin,Huo Jingqian
Nanobodies specifically recognizing atrazine were successfully obtained through immunization of alpacas and phage display libraries, and an indirect competitive immunoassay (ic-ELISA) based on nanobodies was constructed with an IC 50 of 0.062 μg/mL and a minimum detection limit of 0.01 μg/mL. An Au@PtNPs with excellent colorimetric performance, high catalytic activity, and high stability was synthesized. The combination of Au@PtNPs and nanobodies led to the development of a direct competitive immunoassay (dc-ELISA) with higher specificity, convenience, lower cost and an IC 50 value of 0.032 μg/mL which was lower than the ic-ELISA. Based on the Au@PtNPs and nanobodies, a novel LFIA model for qualitative and semi-quantitative detection of atrazine was constructed with a visual detection limit of 5 ng/mL. The present study is of great significance for the construction of atrazine immunoassay combining nanobodies and nanoenzymes, which provides a sensitive, specific, simple and reliable method for the detection of atrazine in the environment.
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研究领域:
分析化学
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免疫学
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纳米科技
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环境科学
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生物化学
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产品关联
DOI
PubMed
高影响因子
FOOD AND AGRICULTURAL IMMUNOLOGY
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2024
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IF 1.7
Jiaxin Wang,Wenhua Shang,Shibei Shao,Jialin Zhang,Qiuyang Wu,Min He,Licai Ma,Jianzhong Shen,Zhanhui Wang,Kai Wen
Paxilline (PAX) is a tremorgenic mycotoxin that is thought to be associated with various syndromes in animals. To facilitate the determination of this toxin, a hapten was chemically synthesized and then respectively conjugated to Bovine Thyroglobulin (BTG) and Bovine serum albumin (BSA) for antibody production. Monoclonal antibody (mAb) 4F8 was finally obtained and used to establish an accurate and sensitive indirect competitive ELISA (icELISA). Various parameters that could impact the performance of icELISA were optimized, including the ion concentration, pH value and organic solvent. Under optimal condition, this assay exhibits an IC50 value as low as 0.84 µg L−1. The limit of detection (LOD) of the established icELISA method for PAX was 0.03–0.3 µg L−1. In conclusion, this study provided a rapid, sensitive, and high-throughput immunoassay for the detection of PAX in grains.
研究领域:
免疫学
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食品科学
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毒理学
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DOI
高影响因子
BIOSENSORS & BIOELECTRONICS
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2024
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IF 10.7
Wenjing Xue,Li Wang,Kexin Yi,Lingyu Sun,Haozhen Ren,Feika Bian
Hepatocellular carcinoma (HCC), as one of the most lethal cancers, significantly impacts human health. Attempts in this area tends to develop novel technologies with sensitive and multiplexed detection properties for early diagnosis. Here, we present novel hydrogel photonic crystal (PhC) barcodes with tyramine deposition amplified enzyme-linked immunosorbent assay (ELISA) for highly sensitive and multiplexed HCC biomarker screening. Because of the abundant amino groups of acrylic acid (AA) component, the constructed hydrogel PhC barcodes with inverse opal structure could facilitate the loading of antibody probes for subsequent detection of tumor markers. By integrating tyramine deposition amplified ELISA on the barcode, the detection signal of tumor markers has been enhanced. Based on these features, it is demonstrated that the hydrogel PhC barcodes with tyramine deposition amplified ELISA could realize highly sensitive and multiplexed detection of HCC-related biomarkers. It was found that this method is flexible, sensitive and accurate, suitable for multivariate analysis of low abundance tumor markers and future cancer diagnosis. These features make the newly developed PhC barcodes an innovation platform, which possesses tremendous potential for practical application of low abundance targets.
研究领域:
生物医学工程
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癌症研究
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分子诊断
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DOI
PubMed
高影响因子
ANALYTICAL AND BIOANALYTICAL CHEMISTRY
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2015
·Vol. 408
·期 4
·1015-1022
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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
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生物医学
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DOI
PubMed
高影响因子
Analytical Methods
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2014
·Vol. 6
·期 13
·4742-4745
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IF 1.938
Liqiang Liu,Dezhao Kong,Changrui Xing,Xun Zhang,Hua Kuang,Chuanlai Xu
Lactoferrin (LF) content in infant milk powder has been strictly regulated by many governments and there is a need for convenient and reliable assays. Using hybridoma techniques, fourteen monoclonal antibodies (mAbs) active against LF were prepared. Two antibodies (mAb2 and mAb3), recognizing spatially distant epitopes of LF, were selected to establish a sandwich enzyme-linked immunosorbent assay (ELISA). A solution of mAb3 (1 μg mL−1) was coated onto micro-titer plates for LF capture while mAb2 labeled with horseradish peroxidase (2.2 μg mL−1) was used as a detection antibody. Under optimized conditions, the proposed sandwich ELISA was evaluated, and linearly responded to LF standards in a range of 5–600 ng mL−1 and the limit of detection was 3.23 ng mL−1. Lactoferrin samples were able to be determined after simple dilution, and the recovery in fortified milk powder averaged between 98% and 109%. The developed assay showed both high specificity (with no obvious cross-reactivity with related proteins) and reproducibility (the coefficient of variation ranged from 4.5% to 7.1%), indicating the utility of this sandwich ELISA in LF monitoring.
研究领域:
免疫学
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食品科学
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分析化学
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DOI
高影响因子
FOOD AND AGRICULTURAL IMMUNOLOGY
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2015
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IF 0.986
Naifeng Xu,Liguang Xu,Wei Ma,Hua Kuang,Chuanlai Xu
In this study, a highly sensitive monoclonal antibody (McAb) against chloramphenicol (CAP) was successfully raised and characterised by indirect competitive enzyme-linked immunosorbent assay (ic-ELISA). Three antigens coupled with different proteins were synthesised and determined by the UV-vis method. After five rounds of immunisation, antisera were collected from six Balb/C mice and screened by ic-ELISA. The standard inhibition curve of one CAP McAb (labelled 6F11) showed the best sensitivity to CAP after optimisation of the principal working conditions. The half-maximal inhibition concentration (IC50) was calculated to be 0.01 ng/mL and the linear working range was from 0.0028 ng/mL to 0.040 ng/mL with a correlation coefficient of 0.996. This McAb showed high specificity, with negligible cross-reactivities detected towards CAP analogues (florfinicol, florfinicol amine, thiamphenicol). The recovery rate was 98.9–106.8% while the coefficient of variation was 4.45–6.05%. The IC50 value and other parameters obtained for this CAP McAb suggest that it is likely to have a promising future in further applications in combination with different analytical techniques.
研究领域:
免疫学
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分析化学
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产品关联
DOI
高影响因子
Foods
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2021
·Vol. 10
·期 10
·2483
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IF 4.35
Wenzhi Tang,Yangchun Qi,Zhonghong Li
Food analysis based on the enzyme-linked immunosorbent assay (ELISA) is simple, sensitive and rapid, but requires a costly colorimetric instrument. The aim of this work was to develop a portable, low-cost and user-friendly colorimetric instrument for colorimetric ELISA and aflatoxin B1(AFB1) detection. The principle of the developed instrument was employing a light-emitting diode to generate the signal light and using a light-dependent resistor to measure the signal light absorbed by the oxidized 3,3′,5,5′-tetramethyl benzidine. The absorption spectra revealed that the solution absorbed signal light more strongly after reaction with H2SO4, and blue light would be favorably absorbed. Evaluations on the stability and accuracy of the instrument and interference from ambient light showed that the fabricated instrument was stable, accurate, capable of quantitative detection and insensitive to ambient light changes. In addition, this instrument is user-friendly since it could calculate and report the final amount of AFB1to the operator. Measurements of maize and peanuts showed that the instrument provided as accurate results as the professional equipment. With the low fabrication cost (about RMB 129 or USD 20), portability, and user-friendliness, this instrument presents attractive potential in the rapid detection of AFB1.Keywords:ELISA;colorimetric;food safety;mycotoxin;rapid detection
研究领域:
食品科学
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分析化学
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仪器仪表
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产品关联
DOI
PubMed
高影响因子
APPLIED MICROBIOLOGY AND BIOTECHNOLOGY
·
2026
·Vol. 110
·期 1
·156
·
IF 4.7
Xu Haoyue,Zhu Xueliang,Li Qian,Liu Yuxi,Wang Qi,Meng Xuelian
Peste des petits ruminants (PPR) is an acute, highly contagious viral disease caused by Peste des petits ruminants virus (PPRV). PPR has inflicted severe economic losses on livestock industries across numerous countries in Africa, Asia, and Europe, posing a significant threat to the survival of endangered wild ruminant species. In this study, we performed prokaryotic expression of the PPRV nucleocapsid (N) protein and prepared rabbit-derived polyclonal antibodies (pAbs). We also identified the characteristics of two mouse-derived monoclonal antibodies (mAbs) against the PPRV N protein. The selected mAb was labeled with horseradish peroxidase (HRP). A double antibody sandwich enzyme-linked immunosorbent assay (DAS-ELISA) was established to detect PPRV antigen using rabbit-derived pAbs and HRP-conjugated mAbs. The threshold for the DAS-ELISA was determined to be 0.227. The DAS-ELISA demonstrated minimum detection limits of 0.195 μg/mL for the recombinant PPRV N protein and 250 50% tissue culture infectious dose (TCID 50 ) for PPRV. Of note, the assay exhibited no cross-reactivity with Capripoxvirus (CaPV), Orf virus (ORFV), Lumpy skin disease virus (LSDV), Foot-and-mouth disease virus (FMDV), or Parainfluenza virus 5 (PIV5). The repeatability of the DAS-ELISA was validated with both intra-assay and inter-assay coefficients of variation (CV%) being less than 9%, indicating good repeatability. Compared to the results obtained using the commercial ID Vet kit (ID-Vet, Montpellier, France) for PPRV antigen detection on the 240 tissue samples, the DAS-ELISA showed a concordance rate of 93.75% and a kappa value of 0.86, demonstrating the reliability of the developed method. The results showed that the DAS-ELISA established in this study is a reliable diagnostic method for the PPRV detection. Key points • Polyclonal and monoclonal antibodies against PPRV N protein were prepared. • A DAS-ELISA based on mAbs and pAbs for detecting virus infection was developed. • The established DAS-ELISA exhibits excellent sensitivity and specificity.
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DOI
高影响因子
BIOSENSORS & BIOELECTRONICS
·
2016
·
IF 7.476
Junlin Wen,Junhua Chen,Li Zhuang,Shungui Zhou
An ultrasensitive and high-throughput nucleic acid detection system, termed as strand displacement reaction-enzyme linked immunosorbent assay (SDR-ELISA), has been developed on the basis of antibody-like DNA nanostructures . Three digoxigenin or biotin modified hairpin probes are utilized to construct antibody-like DNA nanostructures that feature affinity toward streptavidin and anti-digoxigenin antibody via isothermal target-triggered SDR amplification. These antibody-like nanostructures have been employed to conjugate horseradish-peroxidase-labeled anti-digoxigenin antibody with streptavidin that is immobilized on microliter plate wells for enzyme-linked colorimetric assay . The resulting SDR-ELISA system is ultrasensitive for target DNA with a low detection limit of 5 fM. Moreover, the SDR-ELISA system is capable of discriminating DNA sequences with single base mutations, and do so in a high-throughput manner by detection and quantification of up to 96 or 384 DNA samples in a single shot. This detection system is further applied to detect other DNA targets such as Shewanella oneidensis specific DNA sequence, which indicates the generality of proposed SDR-ELISA system. The integration of SDR amplification and convenient ELISA technique advances an intelligent strategy for ultrasensitive and high-throughput nucleic acid detection, which may be amenable for direct visual detection and quantification using an accompanying quantitative color chart.
研究领域:
生物化学
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分子生物学
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医学诊断
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产品关联
DOI
PubMed
高影响因子
Chemosensors
·
2023
·Vol. 11
·期 8
·453
·
IF 4.2
Kai Mao,Lili Tian,Yujie Luo,Qian Li,Xi Chen,Lei Zhan,Yuanfang Li,Chengzhi Huang,Shujun Zhen
Enzyme-linked immunosorbent assay (ELISA) is one of the most commonly used method for the detection of staphylococcal enterotoxin B (SEB), the main protein toxin causing staphylococcal food poisoning. However, the traditional ELISA reaction needs to be stopped by sulfuric acid to obtain stable colorimetric signal, and it is easily influenced by a colored sample. In order to address this problem, a new ELISA method using zeolite imidazolate skeleton-8 metal-organic framework (ZIF-8 MOF) as a light scattering (LS) reporter for SEB detection was developed in this work. ZIF-8 MOF has the characteristics of high porosity, large specific surface area, clear pore structure, and adjustable size, which is one of the most representative MOFs constructed from Zn2+and 2-methylimidazole (2-mIM). The 2-mIM ligand of ZIF-8 exhibited antioxidant activity and can strongly react with H2O2, which could destroy the structure of ZIF-8, resulting in the obvious decrease in LS intensity. We combined this specific reaction with the sandwich immune reaction to construct the LS ELISA method for the successful detection of SEB. This method is more reliable than commercial tests kits for the detection of colored samples, and it is simple, sensitive, and selective, and has great potential in the detection of other toxins by simply changing the corresponding recognition units.Keywords:ELISA;light scattering;metal organic framework;ZIF-8;staphylococcal enterotoxin B
研究领域:
生物化学
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食品安全检测
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免疫学
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产品关联
DOI
高影响因子
危险品化学品经营许可证（带存储）