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
JOURNAL OF AGRICULTURAL AND FOOD CHEMISTRY
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2023
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IF 6.1
Mingyang Li,Xiaoyu Zhang,Yiwen Zhu,Xiancheng Zhang,Zhiyong Cui,Ninglong Zhang,Yue Sun,Zhiying Yang,Wenli Wang,Cunli Wang,Yin Zhang,Yuan Liu,Guangyan Qing
Umami peptides are small molecular weight oligopeptides that play a role in umami taste attributes. However, the identification of umami peptides is easily limited by environmental conditions, and the abundant source and high chromatographic separation efficiency remain difficult. Herein, we report a robust strategy based on a phage random linear heptapeptide library that targets the T1R1-Venus flytrap domain (T1R1-VFT). Two candidate peptides (MTLERPW and MNLHLSF) were readily identified with high affinity for T1R1-VFT binding (KD of MW-7 and MF-7 were 790 and 630 nM, respectively). The two peptides exhibited umami taste and significantly enhanced the umami intensity when added to the monosodium glutamate solution. Overall, this strategy shows that umami peptides could be developed via phage display technology for the first time. The phage display platform has a promising application to discover other taste peptides with affinity for taste receptors of interest and has more room for improvement in the future.
研究领域:
生物化学
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分子生物学
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食品科学
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味觉感知
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DOI
PubMed
高影响因子
ANTIVIRAL RESEARCH
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2014
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IF 3.434
Long Yin,Yuzi Luo,Bo Liang,Fei Wang,Min Du,Valery A. Petrenko,Hua-Ji Qiu,Aihua Liu
Classical swine fever (CSF) is a devastating infectious disease caused by classical swine fever virus (CSFV). The screening of CSFV-specific ligands is of great significance for diagnosis and treatment of CSF. Affinity selection from random peptide libraries is an efficient approach to discover ligands with high stability and specificity. Here, we screened phage ligands for the CSFV E2 protein from f8/8 landscape phage display library by biopanning and obtained four phage clones specific for the E2 protein of CSFV. Viral blocking assays indicated that the phage clone displaying the octapeptide sequence DRATSSNA remarkably inhibited the CSFV replication in PK-15 cells at a titer of 10 10 transduction units, as evidenced by significantly decreased viral RNA copies and viral titers. The phage-displayed E2-binding peptides have the potential to be developed as antivirals for CSF.
研究领域:
病毒学
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分子生物学
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医学
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DOI
PubMed
高影响因子
FOOD CHEMISTRY
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2025
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IF 9.8
Fengchun Zhao,Guanqun Wang,Xinrui Cao,Wenxu Song,Min Guo,Huimin Wang,Zhengyou Yang
Zearalenone (ZEN) is a widely distributed mycotoxin with potent estrogenic activity. Detecting ZEN is crucial for assessing its potential health risks. This study developed a highly sensitive non-competitive magnetic phage anti-immunocomplex immunoassay (Nc-MPHAIA) for ZEN detection, utilizing the anti-ZEN single-chain variable fragment (ScFv) and anti-immunocomplex peptide (AIcP), both of which were screened using phage display technology. The method exhibited a limit of detection of 0.005 ng/mL, which was 112 times lower than that of the conventional competitive immunoassay using the same ScFv and the chemosynthetic antigen ZEN-OVA. Validation studies demonstrated excellent recovery rates in spiked corn samples (intra-assay: 84.04–109.95 %; inter-assay: 89.70–99.59 %). Additionally, the Nc-MPHAIA results for ZEN in natural samples were consistent with those obtained via high-performance liquid chromatography, which demonstrated good reliability of the established assay.
研究领域:
食品安全
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免疫学
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分子生物学
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产品关联
DOI
PubMed
高影响因子
NEW JOURNAL OF CHEMISTRY
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2022
·Vol. 46
·期 21
·10299-10307
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IF 3.925
Xiao-Qin Yang,Li-Wen Bai,Yu Chen,Yue-Xiao Lin,Hua Xiang,Ting-Ting Xiang,Shuang-Xing Zhu,Li Zhou,Kai Li,Xinxiang Lei
Polypeptides provide unique possibilities for the development of efficient and selective fluorescent probes for different biomarkers because of their highly modular nature, the feasibility of chemical synthesis and fluorescence modification, and biomolecular recognition potential. Herein, a bacteriophage (phage) display was utilized to screen affinity peptides against dihydrofolate reductase. On this basis, positive peptide DHFR-Y1(NH2-WSLGYTG-COOH) was obtained utilizing solid-phase peptide synthesis. Furthermore, fluorescein FAM was used to chemically modify the N-terminus and C-terminus of the peptide, respectively, to generate N-terminal fluorescent peptide DHFR-Yn (Flu-βA-WSLGYTG-COOH) and C-terminal fluorescent peptide DHFR-Yc (NH2-WSLGYTL(Flu)-COOH). In the subsequent experiment, multiple in vitro detection methods (including DSF, STD-NMR, BLI) indicated that the binding ability of affinity peptide DHFR-Y1 (KD = 3.57 μM) is significant to DHFR, in contrast to the stronger noncovalent binding ability of C-terminal fluorescent peptide (KD = 0.56 μM) and weak affinity of N-terminal fluorescent peptide (KD = 14.70 μM). The evidence that DHFR-Y1 and DHFR-Yc bind to folic acid binding pocket of DHFR was coming up by the in silico study. The peptide DHFR-Yc is adequate as a fluorescent probe targeting dihydrofolate reductase.
研究领域:
生物化学
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分子生物学
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药物开发
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产品关联
DOI
高影响因子
INTERNATIONAL JOURNAL OF PHARMACEUTICS
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2023
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IF 5.8
Xiaoxu Li,Ximing Pu,Xingming Wang,Juan Wang,Xiaoming Liao,Zhongbin Huang,Guangfu Yin
The obstruction of blood-brain barrier (BBB) and the poor specific targeting are still the major obstacles and challenges of targeted nano-pharmaceutical therapy for glioblastoma (GBM) up to now. It is critical to find appropriate targeting ligands that can effectively mediate the nano-pharmaceuticals to penetrate brain capillary endothelial cells (BCECs) and then specifically bind to glioblastoma cells (GCs). Herein, a dual-targeting ligand for GBM was screened by the combination of phage display peptide library biopanning and affinity-adaptability analysis. Based on the acquisition of sub-library of peptide which exhibited the specific affinity to both BCECs and GCs, a comparison parameter of relative affinity was deliberately introduced to evaluate the relative affinity of candidate peptides to U251-MG cells and bEnd.3 cells. The optimized WTW peptide (sequenced as WTWEYTK) was provided with a high relative affinity (R U/B =2.44), implying that its high affinity to U251-MG cells and moderate affinity to bEnd.3 cells might synergistically promote its receptor-mediated internalization and transport, the dissociation from bEnd.3, and the binding to U251-MG. The results of BBB model trials in vitro showed that the BBB penetration efficiency and GBM accumulation of WTW peptide were significantly higher than those of WSL peptide, GNH peptide, and REF peptide. Results of orthotopic GBM xenograft model assays in vivo also indicated that WTW peptide had successfully penetrated the BBB and improved accumulation in GBM. The screened WTW peptide might be the potential dual-targeting ligand to motivate the advancement of GBM targeted therapy.
研究领域:
癌症生物学
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药理学
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神经科学
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分子生物学
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产品关联
DOI
PubMed
高影响因子
ANALYTICAL CHEMISTRY
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2014
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IF 5.825
Qiaolin Lang,Fei Wang,Long Yin,Mingjun Liu,Valery A. Petrenko,Aihua Liu
Probes against targets can be selected from the landscape phage library f8/8, displaying random octapeptides on the pVIII coat protein of the phage fd-tet and demonstrating many excellent features including multivalency, stability, and high structural homogeneity. Prostate-specific antigen (PSA) is usually determined by immunoassay, by which antibodies are frequently used as the specific probes. Herein we found that more advanced probes against free prostate-specific antigen (f-PSA) can be screened from the landscape phage library. Four phage monoclones were selected and identified by the specificity array. One phage clone displaying the fusion peptide ERNSVSPS showed good specificity and affinity to f-PSA and was used as a PSA capture probe in a sandwich enzyme-linked immunosorbent assay (ELISA) array. An anti-human PSA monoclonal antibody (anti-PSA mAb) was used to recognize the captured antigen, followed by horseradish peroxidase-conjugated antibody (HRP-IgG) and o-phenylenediamine, which were successively added to develop plate color. The ELISA conditions such as effect of blocking agent, coating buffer pH, phage concentration, antigen incubation time, and anti-PSA mAb dilution for phage ELISA were optimized. On the basis of the optimal phage ELISA conditions, the absorbance taken at 492 nm on a microplate reader was linear with f-PSA concentration within 0.825–165 ng/mL with a low limit of detection of 0.16 ng/mL. Thus, the landscape phage is an attractive biomolecular probe in bioanalysis.
研究领域:
分子生物学
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免疫学
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生物技术
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临床诊断
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DOI
PubMed
高影响因子
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