MICROCHEMICAL JOURNAL
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2025
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IF 5.1
Haohan Zhang,Mingyang Wang,Yuxuan Jiang,Zhonghuang Yang,Xin Liu,Mingjun Liu,Wanjian Liu,Aihua Liu
Diabetes mellitus is a global non-communicable disease. Glycated hemoglobin (HbA1c) is the gold standard for assessing long-term blood sugar levels, which is consistent with the average blood sugar concentration and can reflect the patient’s blood sugar control for 2–3 months. The detection of HbA1c is essential for the diagnosis of diabetes and the risk assessment of complications. Traditional HbA1c detection methods have limitations such as significant performance differences, complex technology and high cost. Herein we screened HbA1c-specific peptide by 3 rounds of negative biopanning against hemoglobin and four rounds of conventional panning against HbA1c, by which a specific peptide probe W10 (amino acid sequence: DLRKAGKKRTTS) with high affinity for HbA1c was explored. Then we constructed a label-free electrochemical impedance spectroscopy immunosensor using the W10-displayed phage as biorecognition probe. Under the optimized conditions, the sensor had a linear range of 1–1000 pg/mL and a detection limit of 0.32 pg/mL HbA1c (S/N = 3). The as-proposed sensor demonstrated selectivity against Hb and other blood components and good long-term stability. Clinical validation with 20 human blood samples showed < 7.1 % deviation from HPLC results, enabling direct HbA1c quantification without enzymatic pretreatment. Thus this work establishes a novel, cost-effective platform for diabetes monitoring, which have potential applications in resource-limited settings. This study shows that sensors based on peptide-displaying phages are expected to have potential applications in bioassay for diagnosis and therapy.
研究领域:
医学诊断
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生物技术
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分析化学
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DOI
高影响因子
ACS Applied Bio Materials
·
2020
·
IF 0
Xiangdong Cai,Wenyu Wei,Zhao Liu,Zhongtian Bai,Junqiang Lei,Jianxi Xiao
The development of robust collagen assays is crucial in the diagnosis and treatment of various pathological conditions. Peptide probes composed of the (Gly-Pro-Hyp)n sequences have received extensive attention for their remarkable collagen-targeting capability, which unfortunately has been severely impaired by their high triple helical stability. Herein, we report an efficient strategy to reduce the triple helical propensity of the (Gly-Pro-Hyp)n sequences by electrostatic repulsion. A series of peptides consisting of the (Gly-Pro-Hyp)7 sequence and a number of charged amino acid Asp have been investigated, indicating that the presence of six additional Asp pronouncedly weakened the triple helical stability of peptide probe FAM-PCTP-D6 under physiological conditions (pH 7.4). FAM-PCTP-D6 could be directly applied without any pretreatment to recognize denatured collagen with high selectivity, whereas another dye-labeled peptide probe ROX-PCTP-D6 specifically targeted pathological collagen in various connective tissues of animal disease models and human patients. The inclusion of extra charged natural amino acids has been demonstrated as a convenient approach to create biocompatible collagen-targeting peptide probes with much weaker triple helical stability. Without the need for preheating treatment, these electrostatic repulsion-driven peptide probes provide a handy tool for histopathology staining, showing promising applications in collagen-involved diseases.
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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DOI
PubMed
NEW JOURNAL OF CHEMISTRY
·
2022
·Vol. 46
·期 21
·10299-10307
·
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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DOI
高影响因子
ACS Omega
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2020
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IF 2.87
Wenyu Wei,Dongfang Li,Xiangdong Cai,Zhao Liu,Zhongtian Bai,Jianxi Xiao
The construction of potent peptide probes for selectively detecting denatured collagen is crucial for a variety of widespread diseases. However, all of the denatured collagen-targeting peptide probes found till date primarily utilized the repetitive (Gly-X-Y)n sequences with exclusively imino acids Pro and Hyp in the X and Y positions, which stabilized the triple helical conformation of the peptide probes, resulting in severe obstacles for their clinical applications. A novel series of peptide probes have been constructed by incorporating nonimino acids at the X position of the (GPO)3GXO(GPO)4 sequence, while the X-site residue is varied as Tyr, Phe, Asp, and Ala, respectively. Peptide probes FAM-GYO and FAM-GFO containing aromatic residues Tyr and Phe at the X position showed similarly high binding affinity and tissue-staining efficacy as the well-established peptide probe FAM-GPO, while peptide probes FAM-GDO and FAM-GAO with the corresponding charged residue Asp and the hydrophobic residue Ala indicated much weaker binding affinity and tissue-staining capability. Furthermore, FAM-GYO and FAM-GFO could specifically detect denatured collagen in different types of mouse connective tissues and efficiently stain various human pathological tissues. We have revealed for the first time that the incorporation of nonimino acids, particularly aromatic residues at the X and Y positions of the repetitive (Gly-X-Y)n sequences, may provide a convenient strategy to create novel robust collagen-targeting peptide probes, which have promising diagnostic applications in collagen-involved diseases.
研究领域:
生物化学
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分子生物学
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病理学
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医学诊断
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产品关联
DOI
PubMed
高影响因子
Scientific Reports
·
2017
·Vol. 7
·期 1
·1-12
·
IF 4.259
Liu Liang,Xu Qingqing,Hao Shuai,Chen Yun
MicroRNAs (miRNAs) play a vital role in regulating gene expression and are associated with a variety of cancers, including breast cancer. Their distorted and unique expression is a potential marker in clinical diagnoses and prognoses. Thus, accurate determination of miRNA expression levels is a prerequisite for their applications. However, the assays currently available for miRNA detection typically require pre-enrichment, amplification and labeling steps, and most of the assays are only semi-quantitative. Therefore, we developed a quasi-direct liquid chromatography-tandem mass spectrometry (LC-MS/MS)-based targeted proteomics approach to quantify target miRNA by innovatively converting the miRNA signal into the mass response of a reporter peptide via a covalently immobilized DNA-peptide probe. Specifically, the probe containing the targeted proteomics-selected substrate/reporter peptide, GDR AVQLGVDPFR/AVQLGVDPFR, and the DNA sequence complementary to the target miRNA (i.e., miR-21) was first immobilized on APMTS modified silica nanoparticles using PDITC. After the immobilized probe was recognized and hybridized with the target miRNA, the excess probe was degraded using MBN and followed by a trypsin digestion of the hybrids. The reporter peptide was released and quantified using LC-MS/MS. The obtained LOQ was 5 pM. Finally, the developed assay was used for the quantitative analysis of miR-21 in breast cells and tissue samples.
研究领域:
分子生物学
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癌症研究
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蛋白质组学
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分析化学
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产品关联
DOI
PubMed
高影响因子
ANALYTICA CHIMICA ACTA
·
2025
·
IF 6
Zongmei Zheng,Mingyang Wang,Jinlong Yuan,Junchong Liu,Haipeng Yu,Zhonghuang Yang,Wanjian Liu,Aihua Liu
It is crucial to timely and accurately identify the causative virus for early treatment and urgent prevention. Viral antigen detection can identify those people who are most likely at risk of spreading the disease, but most based on antibodies with limited stability and sensitivity. Peptides offer several advantages over antibodies, such as low cost, smaller size and good stability. The development of electrochemical immunoassay using specific peptide probes have the merits of good sensitivity and selectivity as well as good stability. Herein we report an amperometric immunosensor using peptides as capture probe and recognition probe. The molecular docking suggests that the two peptides of Pi (sequence: NFWISPKLAFALGGGKKKSC) and FK11 (sequence: WFLNDSELISML), bioscreened from phage display, bind to N-terminal domain of SARS-CoV-2 spike protein (SP). The peptide Pi is assembled on MoS 2 @AuNPs modified electrode to capture SARS-CoV-2 SP, which is recognized by peptide FK11-displayed phage to form Pi/SARS-CoV-2 SP/FK11-phage sandwich. Then anti-M13 phage conjugated horseradish peroxidase (HRP) (anti M13-HRP) was introduced to recognize the phage capsid protein pVIII to form M13 phage/anti M13-HRP to enrich thousands of HRP, which can further electrochemically catalyze H 2 O 2 reduction at highly conductive MoS 2 @AuNPs at − 0.35 V. Then amperometric immunosensor was constructed with linear range of 0.1−5000 pg/mL SARS-CoV-2 SP and detection limit of 0.074 pg/mL. The sensor also has good selectivity, batch reproducibility and stability, capable of detecting down to 10 transducing units/mL SARS-CoV-2 pseudoviruses. This work represents the first example of dual-peptide probes based sandwich-type electrochemical immunosensor integrated with dual signal amplification, which may provide a cost-effective assay platform in detecting real SARS-CoV-2 viruses for early diagnosis. The flexible and modular strategy can be extended to develop other type biosensors for a wide range of applications.
研究领域:
生物医学工程
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病毒学
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分析化学
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产品关联
DOI
PubMed
高影响因子
ANALYTICAL CHEMISTRY
·
2015
·
IF 5.636
Feifei Xu,Ting Yang,Yun Chen
The distorted and unique expression of microRNAs (miRNAs) in cancer makes them an attractive source of biomarkers. However, one of prerequisites for the application of miRNAs in clinical practice is to accurately profile their expression. Currently available assays normally require pre-enrichment, amplification, and labeling steps, and most of them are semiquantitative. In this study, we converted the signal of target miR-21 into reporter peptide by a DNA-peptide probe and the reporter peptide was ultimately quantified using LC-MS/MS-based targeted proteomics. Specifically, substrate peptide GDKAVLGVDPFR containing reporter peptide AVLGVDPFR and tryptic cleavage site (lysine at position 3) was first designed, followed by the conjugation with DNA sequence that was complementary to miR-21. The newly formed DNA-peptide probe was then hybridized with miR-21, which was biotinylated and attached to streptavidin agarose in advance. After trypsin digestion, the reporter peptide was released and monitored by a targeted proteomics assay. The obtained limit of quantification (LOQ) was 1 pM, and the detection dynamic range spanned ∼5 orders of magnitude. Using this assay, the developed quasi-targeted proteomics approach was applied to determine miR-21 level in breast cells and tissue samples. Finally, qRT-PCR was also performed for a comparison. This report grafted the strategy of targeted proteomics into miRNA quantification.
研究领域:
癌症生物学
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分子生物学
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蛋白质组学
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分析化学
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产品关联
DOI
PubMed
高影响因子
ADVANCED FUNCTIONAL MATERIALS
·
2020
·Vol. 30
·期 42
·2004532
·
IF 16.836
Xiangdong Cai,Wenyu Wei,Yihui Bi,Zhao Liu,Zhongtian Bai,Junqiang Lei,Jianxi Xiao
The efficient detection of pathological collagen plays an essential role in the diagnosis, prognosis, and treatment of various critical diseases such as arthritis and tumors. Peptide probes with the (Gly-Pro-Hyp)n sequences have been recently discovered to recognize pathological collagen; however, their staining efficacy severely suffers from their strong trimerizing tendency. An intrinsically nontrimerizing peptide probe F-GOP-10 has been constructed for the first time to specifically target pathological collagen. F-GOP-10 consists of the repetitive (Gly-Hyp-Pro)n sequences, and displays the single stranded conformation at 0 °C. F-GOP-10 has been demonstrated to selectively recognize pathological, but not intact collagen in various mouse connective tissues, and it has been successfully applied to detect pathological collagen in osteoarthritis, liver fibrosis, and various cancer tissues. Compared with the common Masson's trichrome staining method, F-GOP-10 displays unique selectivity to only bind to pathological collagen without the interference of intact collagen. Notably, peptide probes with different lengths of the Gly-Hyp-Pro triplets all maintain the single stranded conformation, demonstrating their intrinsically non-trimerizing nature. This pure-peptide, pure monomer probe allows the accurate concentration determination and the elimination of harmful pretreatment, which has great potential in histopathology staining and other clinical applications.
研究领域:
医学诊断
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病理学
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分子生物学
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生物医学工程
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产品关联
DOI
高影响因子
Journal of Materials Chemistry B
·
2019
·Vol. 7
·期 48
·7676-7682
·
IF 5.047
Xiuxia Sun,Linyan Yao,Caihong Fu,Liting Luo,Jie Wang,Jianxi Xiao
Collagen with a single amino acid substitution is the main cause of a plethora of heritable disorders such as Osteogenesis Imperfecta and Ehlers-Danlos syndrome. Though significant advances have been achieved in the development of protein assays, it remains very challenging to distinguish a protein with a single amino acid mutation from the wild-type protein. A novel fluorescent self-quenching assay has been constructed to detect target collagen peptides with a single amino acid mutation using two probe peptides. The hybridization of the probe peptide and the natural target collagen peptide results in a complete heterotrimer and strong fluorescence, whereas the mixture of the probe peptide and the mutation collagen sequences leads to a partial homotrimer and pronounced fluorescence self-quenching. The extent of fluorescence quenching is dependent on the identity of the residue replacing Gly following the order of Ala < Ser < Arg, while the Gly–Ala mutation causes the mildest fluorescence loss. The probe peptide-based fluorescence self-quenching assay facilitates specific detection of the target collagen sequence with a single Gly mutation at the nM level. The simultaneous utilization of both probe peptides enables efficient discrimination between different mutation peptides. To our knowledge, our work may be the first report of a robust analytical assay that can identify collagen fragments with single amino acid mutation, which will greatly contribute to deciphering the molecular mechanism of Osteogenesis Imperfecta as well as developing novel diagnostic strategies.
研究领域:
生物化学
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分子生物学
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分析化学
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医学诊断
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产品关联
DOI
PubMed
高影响因子
ANALYTICAL CHEMISTRY
·
2023
·
IF 7.4
Qishu Jiao,Yaxin Zheng,Shicheng Pei,Xuan Luo,Xiaoxing Wu,Keming Xu,Wenying Zhong
Noninvasive monitoring of chymotrypsin-like (ChT-L) activity of proteasomes is of great significance for the diagnosis and prognosis of various cancers. However, commercially available proteasome probes usually lack adequate cancer-cell selectivity. To noninvasively monitor ChT-L activity of proteasomes in living cells, we rationally designed a cascade-activated AIEgen-peptide probe (abbreviated as TPE-1p), which self-assembled in aqueous solution to exhibit bright fluorescence in response to sequential treatment of alkaline phosphatase (ALP) and ChT-L. Transmission electron microscopy, enzymatic kinetics, and in vitro fluorescence experiments validated that TPE-1p was efficiently dephosphorylated by ALP to generate TPE-1, which was recognized by ChT-L in the proteasome, and transformed to form nanofibers with strong fluorescence signals. Cell imaging experiments revealed that bright blue fluorescence was observed in TPE-1p-treated HeLa cells, whereas NIH3T3 and HepG2 cells showed less fluorescence at the same condition. The enhanced fluorescence signals in HeLa cells were attributed to the high activities of endogenous ALP and ChT-L. Moreover, TPE-1p was utilized to noninvasively assess the inhibition efficiency of a ChT-L inhibitor (bortezomib, abbreviated as Btz) in HeLa cells. Significant correlation was found between the fluorescence signals of TPE and the viabilities of Btz-treated cells in concentration ranges from 0 to 1 μM, indicating that TPE-1p could be employed to predict the activity of ChT-L inhibitors. The design of the cascade-activated AIEgen-peptide probe provides a viable approach for noninvasively monitoring the ChT-L activity of proteasomes in living cells, which facilitates high-throughput screening of ChT-L inhibitors in cancer therapy.
研究领域:
癌症生物学
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分子生物学
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生物医学工程
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产品关联
DOI
PubMed
高影响因子
JOURNAL OF NANOBIOTECHNOLOGY
·
2025
·
IF 12.6
Huang Xiaobin,Yi Yakun,Zhou Ke,Liu Lei,Wu Hai-Chen
MicroRNAs (miRNAs) play crucial roles in the pathogenesis of diverse diseases; however, their extremely low abundance in complex biological systems makes highly sensitive and specific multiplexed detection particularly challenging. Here, we report an enzyme-assisted nanopore sensor that enables simultaneous and ultrasensitive detection of multiple miRNAs. Capture DNA (cDNA) immobilized on streptavidin-coated magnetic beads specifically hybridizes with target miRNAs, opening the cDNA hairpin and facilitating the binding of alkaline phosphatase (ALP)-labeled detection DNA to form stable complexes. ALP catalyzes the dephosphorylation of designed phosphorylated peptide probes, inducing conformational and charge changes that yield distinct current blockade signatures in the presence of cucurbit [ 7 ]uril within the nanopore. This strategy reduces background interference via magnetic separation and amplifies detection signals through enzymatic reactions, thereby markedly improving sensitivity for low-abundance miRNAs. The sensor achieves a broad dynamic range from 10.0 fM to 10.0 nM across three target miRNAs, with femtomolar-level limits of detection. By integrating molecular separation, enzymatic amplification, and nanopore sensing, this platform provides a robust approach for precise miRNA analysis, offering strong potential for early disease diagnosis.
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产品关联
DOI
PubMed
高影响因子
TALANTA
·
2024
·
IF 5.6
Jean Claude Munyemana,Xiuxia Sun,Lu Li,Chunxia Zhang,Eskandar Qaed,Jianxi Xiao
ABSTRACT Monitoring collagen denaturation is crucial for diagnosing collagen-related diseases such as tumors and fibrosis. Herein, we have developed specific probes to detect denatured collagen (d-Col) and collagen I (Col I), utilizing peptide probes with sequences (GOP)10 and HVWMQAP, targeting at d-Col and Col I, respectively. These peptides were conjugated with 1,10-phenanthroline-5-carboxylic Acid (Phen), forming Phen-Ahx-(GOP)10 and Phen-Ahx-HVWMQAP. Phen acts as both an antenna sensitizer and a chelator, coordinating with Terbium (III) and Europium (III) ions via its nitrogen atom, facilitating fluorescent emission in green and red, respectively. The investigation demonstrated that Tb 3+ interacts with three (GOP) 10 peptide units through Phen, while Eu 3+ connects with four units of Ahx-HVWMQAP peptides. Additionally, it is important to note that the structure of the peptides remains unchanged after chelating with the lanthanide ions, maintaining their integrity throughout the process. These probes have effectively demonstrated their ability to bind to specific collagen types with selectivity, enabling accurate identification of their presence. The excellent binding of these probes is due to the branched structure of the formed lanthanide-peptide complexes. A dose-dependent linear association was observed in the binding of Eu-[Phen-Ahx-HVWMQAP] 4 to Col I, with concentration levels ranging from 0.5 to 100 μM and a minimal detectable concentration of 0.113 μM. We anticipate that our developed probes will improve understanding of collagen remodelling and provide opportunities for the diagnosis of collagen- associated diseases.
研究领域:
生物材料
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生物医学工程
·
分子生物学
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分析化学
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材料科学
✓ 已验证
DOI
PubMed
高影响因子
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