ELECTROCHIMICA ACTA
·
2022
·
IF 7.336
Guiyin Li,Bo Wang,Le Zhao,Xiaohang Shi,Guanxiong Wu,Wei Chen,Lixian Sun,Jintao Liang,Zhide Zhou
Glypican-3 (GPC3) protein is considered as an effective and specific biomarker for the accurate diagnosis of hepatocellular carcinoma (HCC), and there is a critical demand to construct strategies with shorter time, lower cost noise and higher sensitivity. In this study, a biosensor for label-free detection of GPC3 was prepared by modifying GPC3 aptamer (GPC3 Apt ) on the surface of Si-based light addressable potentiometric sensor (LAPS) chip modified by reduced graphene oxide/polyetherimide/gold nanoparticles (rGO/PEI/AuNPs). Among them, GPC3 Apt with a low equilibrium dissociation constant was used as a biorecognition molecule to recognize and capture GPC3. The rGO/PEI/AuNPs decorated on the LAPS surface could enlarge the surface area for GPC3 Apt immobilization and provide more active sites for GPC3 Apt to bind effectively with the target GPC3. When a bias voltage was provided to the LAPS system, the forming GPC3-GPC3 Apt complexs on the sensing interface of LAPS would cause the potential of the sensitive membrane surface to change and generate a corresponding offset response in photocurrent-voltage (I-V) curves. The potential shift and GPC3 concentration showed a good linear range from 0.1 to 100.0 μg/mL with a detection limit of 40.0 ng/mL (S/ N = 3) and a sensitivity of 1.33638 mV/μg/mL. In addition, the LAPS detection system also has good specificity and stability, which can be used for the detection of GPC3 in spiked serum samples.
研究领域:
生物传感器
·
癌症诊断
·
纳米技术
·
分析化学
·
生物医学工程
✓ 已验证
DOI
高影响因子
IEEE SENSORS JOURNAL
·
2026
·
IF 4.5
Lingling Fan,Cunhong Cen,Yuru Wu,Xiaohua Deng,Yuxuan Shi,Yuxing Huang,Xiaohong Tan,Shidong Li,Guiyin Li
Hepatocellular carcinoma (HCC) remains one of the most lethal malignancies worldwide, underscoring the urgent demand for early diagnostic strategies to improve patient survival rates. Glypican-3 (GPC3), a highly specific biomarker of HCC, has emerged as a promising target for early detection. Herein, we developed an innovative magnetic colorimetric aptasensor using ferrosoferric oxide@carbon quantum dots (Fe3O4@CQDs) nanocomposites. The Fe3O4@CQDs nanocomposites combine the magnetic properties of Fe3O4 with the unique optical characteristics of CQDs, exhibiting intrinsic peroxidase‑like activity. The animo-modified GPC3 aptamer (GPC3Apt) was covalently conjugated onto the Fe3O4@CQDs nanocomposites surface via EDC/NHS chemistry to form the recognition probe (Fe3O4@CQDs-GPC3Apt). Upon specific binding between the aptamer and GPC3, the resulting complexes (Fe3O4@CQDs-GPC3Apt-GPC3) modulate the catalytic oxidation of o-phenylenediamine (OPD) by H2O2, generating a yellow oxOPD product whose absorbance intensity correlated with GPC3 concentration. The proposed colorimetric aptasensor demonstrated a detection limit of 3.16 ng/mL and a linear range of 10–200 ng/mL, with high specificity against interfering proteins. It was successfully validated in human serum samples, yielding recoveries between 99.27% and 109.33%. Owing to its operational simplicity and cost‑effectiveness, this platform offers transformative potential for point-of-care HCC screening. Beyond advancing GPC3 detection, the work provides a flexible design paradigm that can be adapted for assays targeting other disease biomarkers.
✓ 已验证
DOI
高影响因子
MICROCHIMICA ACTA
·
2020
·Vol. 187
·期 5
·1-11
·
IF 6.232
Zhou Zhide,Zhao Le,Li Wenzhan,Chen Min,Feng Huafu,Shi Xiaohang,Liang Jintao,Li Guiyin
A Glypican-3 (GPC3) electrochemical aptamer nanobiosensor based on hemin/graphene nanohybrids (HGNs) peroxidase-like catalytic silver deposition and GPC3 aptamer has been constructed for the determination of GPC3. The HGNs were prepared by an one-step reduction method. Fourier transform infrared spectroscopy (FT-IR), ultraviolet spectroscopy (UV-vis), and transmission electron microscopy (TEM) were used to study the structure and morphological characteristics of the HGNs. The GPC3 electrochemical aptamer nanobiosensor was constructed using HGNs-aptamer (HGNs-Apt) as the signal probe and GPC3 aptamer as the capture probe. With the help of the catalytic action of peroxidase-like properties of HGNs, H 2 O 2 reduces the silver (Ag) ions in solution to metallic Ag, which deposit on the surface of the electrode. The amount of deposited Ag, which was derived from the amount of GPC3, was quantified by differential pulse voltammetry (DPV). Under optimal conditions, the current response of Ag had a good positive correlation with the GPC3 concentration in the range 10.0–100.0 μg mL −1 with a correlation coefficient of 0.9958. The detection limit was 3.16 μg mL −1 at a signal-to-noise ratio of 3, and the sensitivity was calculated to be 0.807 μA μM −1 cm −2 . The method is validated by analyzing spiked human serum samples with good recovery ranging from 101 to 122%. In addition, the GPC3 electrochemical aptamer nanobiosensor has acceptable selectivity, stability, and reproducibility.
A Glypican-3 electrochemical aptamer nanobiosensor based on hemin/graphene nanohybrids (HGNs) peroxidase-like catalytic silver deposition and GPC3 aptamer has been constructed for the determination of GPC3. The electrochemical aptamer nanobiosensor exhibits high selectivity, acceptance reproducibility, and good recovery performances.
研究领域:
生物传感器
·
分析化学
·
纳米技术
·
癌症诊断
✓ 已验证
产品关联
DOI
PubMed
高影响因子
Nature Communications
·
2024
·Vol. 15
·期 1
·1-20
·
IF 14.7
Liu Jinhu,Zhao Huajun,Gao Tong,Huang Xinyan,Liu Shujun,Liu Meichen,Mu Weiwei,Liang Shuang,Fu Shunli,Yuan Shijun,Yang Qinglin,Gu Panpan,Li Nan,Ma Qingping,Liu Jie,Zhang Xinke,Zhang Na,Liu Yongjun
Cytotherapy is a strategy to deliver modified cells to a diseased tissue, but targeting solid tumours remains challenging. Here we design macrophages, harbouring a surface glypican-3-targeting peptide and carrying a cargo to combat solid tumours. The anchored targeting peptide facilitates tumour cell recognition by the engineered macrophages, thus enhancing specific targeting and phagocytosis of tumour cells expressing glypican-3. These macrophages carry a cargo of the TLR7/TLR8 agonist R848 and INCB024360, a selective indoleamine 2,3-dioxygenase 1 (IDO1) inhibitor, wrapped in C16-ceramide-fused outer membrane vesicles (OMV) of Escherichia coli origin (RILO). The OMVs facilitate internalization through caveolin-mediated endocytosis, and to maintain a suitable nanostructure, C16-ceramide induces membrane invagination and exosome generation, leading to the release of cargo-packed RILOs through exosomes. RILO-loaded macrophages exert therapeutic efficacy in mice bearing H22 hepatocellular carcinomas, which express high levels of glypican-3. Overall, we lay down the proof of principle for a cytotherapeutic strategy to target solid tumours and could complement conventional treatment.
研究领域:
肿瘤学
·
免疫治疗
·
药理学
·
细胞生物学
✓ 已验证
DOI
PubMed
高影响因子
BIOELECTROCHEMISTRY
·
2025
·
IF 4.5
Xuyang Chen,Lingling Fan,Kaipeng Su,Shuwei Wen,Xiaohong Tan,Yuge Liu,Xiaohua Deng,Xiangming Li,Yong Huang,Guiyin Li
Hepatocellular carcinoma (HCC) is a high-mortality malignancy that urgently requires sensitive early-stage biomarkers. Glypican-3 (GPC3), a highly specific protein biomarker for HCC, calls for efficient detection strategies. Herein, we developed a label-free electrochemical aptasensor based on nitrogen-doped reduced graphene oxide-ferrocene-polyaniline (NRGO-Fc-PANI) nanocomposites and GPC3 aptamer for GPC3 detection. The NRGO-Fc-PANI combined the large high surface area and conductivity of NRGO, high electrical conductivity and stability of PANI, and the good redox properties of Fc, which functed as in situ electrochemical redox signal indicator to monitor the electrochemical changes. Upon target binding, the formation of GPC3-aptamer complex fell off from the electrode, exposing the conductive NRGO-Fc-PANI layer and enhancing the differential pulse voltammetry (DPV) response. The aptasensor exhibited a linear detection range of 10.0–100.0 ng·mL −1 (R 2 = 0.996) and a low limit of detection (LOD) of 2.88 ng·mL −1 . It demonstrated excellent selectivity toward interfering proteins, retained 70.8 % of its initial signal after 11 days of storage, and attained a recovery range of 97.0–109.5 % in human serum samples. By eliminating the need for enzymatic or fluorescent reporters, the aptasensor reduced reagent costs and operational complexity, presenting a clinically promising tool for the early diagnosis of HCC.
研究领域:
肿瘤学
·
生物传感
·
分子诊断
·
转化医学
·
纳米医学
·
电分析化学
✓ 已验证
产品关联
DOI
PubMed
高影响因子
PROTEIN EXPRESSION AND PURIFICATION
·
2022
·
IF 2.025
Zhiting Lao,Shuanqi Li,Jinhui Liang,Jingyi Su,Xin Gong,Xiping Cui,Suqing Zhao
Glypican-3 (GPC3) has a promise to be the diagnostic biomarker as well as therapeutic target for hepatocellular carcinoma (HCC). Nanobody have the great potential in clinical diagnosis and treatment for their characteristics of small size, high solubility, stability, manipulability, binding advantages, and ease of production. In this study, the recombinant glypican-3-N terminal (GPC3-N) protein was expressed as inclusion body in E. coli BL21(DE3)pLysS cells and then purified, which is then used as the immunogen to construct nanobody phage library. The positive clone (named MF15) was obtained by four rounds of bio-panning, and then transformed into the E. coil TOP10F’ cells to express nanobody protein, with the molecular weight of 19 kDa. Both Western blot and immunofluorescence analysis revealed that bacterially expressed GPC3-N protein is biologically active, and MF15 could specifically recognized native GPC3 expressed in HepG2 cells. The results in this study would provide the technical support for the development of diagnostic kits and antibody drugs targeting GPC3.
研究领域:
分子生物学
·
癌症研究
·
生物技术
✓ 已验证
DOI
PubMed
高影响因子
MICROCHIMICA ACTA
·
2024
·Vol. 191
·期 11
·1-17
·
IF 5.3
Liang Jintao,Liang Jianlu,Xiao Xinkai,Guo Fei,Zhan Yulian,Zhou Xueqing,Zhou Zhide,Li Guiyin
A sandwich-type electrochemical aptasensor for ultrasensitive detection of glypican-3 (GPC3) was constructed using GPC3 aptamer (GPC3 Apt ) labelled reduced graphene oxide-cerium oxide-gold nanoparticles (RGO-CeO 2 -Au NPs) as the signal probe and the same GPC3 Apt as the capture probe. The electrochemical redox properties of CeO 2 (Ce 3+ /Ce 4+ ) in the RGO-CeO 2 -Au NPs indicate the electrochemical signals. When the target GPC3 was present, an “aptamer-protein-aptamer” sandwich structure was formed on the sensing interface due to the specific binding between the protein and aptamers, resulting in an increased electrochemical redox signal detected by differential pulse voltammetry (DPV) technique. Under optimal conditions, the established aptasensor exhibited a logarithmic linear relationship between the current response and GPC3 concentration in the range 0.001–100.0 ng/mL, with a minimum detection limit of 0.74 pg/mL. Using the spike-recovery tests for measurement of the human serum samples, the recovery was from 99.26 to 114.01%, and the RSD range was 3.04 to 5.34%. Furthermore, the sandwich-type electrochemical aptasensor exhibited excellent performance characteristics such as good stability, high specificity, and high sensitivity, demonstrating effective detection of GPC3 in human serum samples and can be used as a clinical detection tool for GPC3. Graphical Abstract
研究领域:
电化学
·
纳米技术
·
生物传感器
·
癌症诊断
✓ 已验证
产品关联
DOI
PubMed
高影响因子
ANALYTICAL AND BIOANALYTICAL CHEMISTRY
·
2022
·Vol. 414
·期 22
·6441-6453
·
IF 4.478
Li Guiyin,Chen Wei,Mi Danhong,Wang Bo,Li HaiMei,Wu Guangxiong,Ding Ping,Liang Jintao,Zhou Zhide
Hepatocellular carcinoma (HCC) is the second leading cause of cancer-related deaths in China. Glypican-3 (GPC3) is a specific antigen related to HCC, which is widely used in clinical detection as a reliable marker of HCC. In this paper, a highly sensitive homogeneous apatasensor was designed for GPC3 detection based on fluorescence resonance energy transfer (FRET) where the GPC3 aptamer labelled gold carbon dots (AuCDs-GPC3 Apt ) are used as a donor and magnetic graphene oxide (Fe 3 O 4 /GO) nanosheets are used as an acceptor. A one-step hydrothermal method was used to synthesize AuCDs to provide sufficient fluorescence. The FRET phenomenon exists between AuCDs-GPC3 Apt and Fe 3 O 4 /GO, which weakens the fluorescence intensity of the whole system. When the target GPC3 is added to the FRET system, the fluorescent AuCDs-GPC3 Apt binds to the GPC3 and forms a folded structure, which leads to AuCDs-GPC3 Apt separation from Fe 3 O 4 /GO nanosheets. The Fe 3 O 4 /GO is then magnetically separated so that the fluorescence of free labelled AuCDs-GPC3 Apt is restored. Under the optimum conditions, the fluorescence recovery rate is linearly correlated with the concentration of GPC3 (5–100 ng·mL −1 ) and the detection limit is 3.01 ng·mL −1 (S/N = 3). This strategy shows recoveries from 98.76 to 101.29% in real human serum samples and provides an immediate and effective detection method for the quantification of GPC3 with great potential applications for early diagnosis of HCC. Graphical abstract A sensitive homogeneous FRET-based apatasensor was designed for GPC3 detection where the AuCDs-GPC3 Apt is a donor and Fe 3 O 4 /GO nanosheets are an acceptor. The GPC3 fluorescent aptasensor combines wider output range with low cost, high specificity, and good anti-interference.
研究领域:
医学诊断
·
癌症研究
·
纳米技术
·
分子生物学
✓ 已验证
产品关联
DOI
PubMed
高影响因子
Journal of Biomedical Nanotechnology
·
2021
·Vol. 17
·期 12
·2444-2454
·
IF 4.099
Li Guiyin,Li HaiMei,Chen Wei,Chen Huijiang,Wu Guanxiong,Tan Mingxiong,Liang Jintao,Zhou Zhide
An electrochemical aptasensor for quantitatively detecting glypican-3 (GPC3) was constructed by combining hemin-reduced graphene oxide-platinum (H-rGO-Pt) nanoparticles (NPs) with reduced graphene oxide-gold (rGO-Au) nanoparticles (NPs). Herein, the rGO-Au NPs deposited onto screen-printed
electrodes resulted in signal amplification due to their large surface areas. Meanwhile, highly conductive H-rGO-Pt NPs acted as a sensing medium that improved electrical conductivity and as an indicator for monitoring peak current for determination. A GPC3 aptamer (GPC3apt) with a low equilibrium
dissociation constant was used as a bio-recognition molecule. GPC3apt specifically captured GPC3 proteins and formed aptamer-GPC3 complexes, which impeded electron transfer and thus hampered the redox signal of hemin in H-rGO-Pt NPs. This developed electrochemical aptasensor showed a linear
response to GPC3 (from 0.001 μg/mL to 10 μg/mL) and had a detection limit of 0.001 μg/mL. This work provides a low-cost and highly sensitive detection with and good recovery for GPC3 and holds great promise for the clinical diagnosis of hepatocellular carcinoma.
研究领域:
生物医学工程
·
肿瘤学
·
纳米技术
✓ 已验证
DOI
PubMed
高影响因子
Biomaterials Science
·
2022
·Vol. 10
·期 11
·2865-2876
·
IF 7.59
Mengyue Pei,Huiyang Li,Yingwei Zhu,Jian Lu,Chuangnian Zhang
Liver cancer is the most common malignant tumor and liver cancer immunotherapy has been one of the research hotspots. To induce antigen-specific antitumor immune responses against liver cancer, we developed antigen and adjuvant co-delivery nanovaccines (APPCs). Polyanionic alginate (ALG) and polycationic polyethyleneimine (PEI) were utilized to co-deliver a glypican-3 peptide antigen and an unmethylated cytosine-phosphate-guanine (CpG) adjuvant by electrostatic interactions. A cellular uptake study confirmed that APPC could promote antigen and adjuvant uptake by dendritic cells (DCs). Importantly, APPC facilitated the endosomal escape of the peptide for antigen delivery into the cytoplasm. In addition, APPC showed significant stimulation of DC maturation in vitro. APPC could also efficiently prime DCs and induce cytotoxic T lymphocyte responses in vivo. The in vitro cell viability assay and the in vivo histocompatibility showed that APPC was non-toxic within the tested concentration. This study demonstrates that the peptide antigen and the CpG adjuvant co-delivery nanovaccine have potential applications in liver cancer immunotherapy.
研究领域:
免疫学
·
肿瘤学
·
分子生物学
·
药剂学
✓ 已验证
产品关联
DOI
PubMed
高影响因子
JOURNAL OF NANOBIOTECHNOLOGY
·
2022
·Vol. 20
·期 1
·1-11
·
IF 9.429
Hsieh Yuan-Chin,Liao Jun-min,Chuang Kuo-Hsiang,Ho Kai-Wen,Hong Shih-Ting,Liu Hui-Ju,Huang Bo-Cheng,Chen I-Ju,Liu Yen-Ling,Wang Jaw-Yuan,Tsai Hsiang-Lin,Su Yu-Cheng,Wang Yen-Tseng,Cheng Tian-Lu
Background Humanization of mouse monoclonal antibodies (mAbs) is crucial for reducing their immunogenicity in humans. However, humanized mAbs often lose their binding affinities. Therefore, an in silico humanization method that can prevent the loss of the binding affinity of mAbs is needed.Methods We developed an in silico V(D)J recombination platform in which we used V(D)J human germline gene sequences to design five humanized candidates of anti-tumor necrosis factor (TNF)-α mAbs (C1–C5) by using different human germline templates. The candidates were subjected to molecular dynamics simulation. In addition, the structural similarities of their complementarity-determining regions (CDRs) to those of original mouse mAbs were estimated to derive the weighted interatomic root mean squared deviation (wRMSDi) value. Subsequently, the correlation of the derived wRMSDi value with the half maximal effective concentration (EC50) and the binding affinity (KD) of the humanized anti-TNF-α candidates was examined. To confirm whether our in silico estimation method can be used for other humanized mAbs, we tested our method using the anti-epidermal growth factor receptor (EGFR) a4.6.1, anti-glypican-3 (GPC3) YP9.1 and anti-α4β1 integrin HP1/2L mAbs.Results The R2 value for the correlation between the wRMSDi and log(EC50) of the recombinant Remicade and those of the humanized anti-TNF-α candidates was 0.901, and the R2 value for the correlation between wRMSDi and log(KD) was 0.9921. The results indicated that our in silico V(D)J recombination platform could predict the binding affinity of humanized candidates and successfully identify the high-affinity humanized anti-TNF-α antibody (Ab) C1 with a binding affinity similar to that of the parental chimeric mAb (5.13 × 10−10). For the anti-EGFR a4.6.1, anti-GPC3 YP9.1, and anti-α4β1 integrin HP1/2L mAbs, the wRMSDi and log(EC50) exhibited strong correlations (R2 = 0.9908, 0.9999, and 0.8907, respectively).Conclusion sOur in silico V(D)J recombination platform can facilitate the development of humanized mAbs with low immunogenicity and high binding affinities. This platform can directly transform numerous mAbs with therapeutic potential to humanized or even human therapeutic Abs for clinical use.Graphical Abstract
研究领域:
免疫学
·
生物信息学
·
分子生物学
·
药物开发
✓ 已验证
产品关联
DOI
PubMed
高影响因子
Advanced Healthcare Materials
·
2023
·Vol. 12
·期 12
·2202424
·
IF 10
Dongdong Guo,Xiaoyong Dai,Kewei Liu,Yuhong Liu,Jiamin Wu,Kun Wang,Shengwei Jiang,Fen Sun,Lijun Wang,Bing Guo,Dongye Yang,Laiqiang Huang
While calcium-overload-mediated therapy (COMT) is a promising but largely untapped therapeutic strategy, combinatory therapy greatly boosts treatment outcomes with integrated merits of different therapies. Herein, a BPQD@CaO 2 -PEG-GPC3Ab nanoplatform is formulated by integrating calcium peroxide (CaO 2 ) and black phosphorus quantum dot (BPQD, photosensitizer) with active-targeting glypican-3 antibody (GPC3Ab), for combinatory photodynamic therapy (PDT) and COMT in response to acidic pH and near-infrared (NIR) light, wherein CaO 2 serves as the reservoir of calcium ions (Ca 2+ ) and hydrogen peroxide (H 2 O 2 ). Navigated by GPC3Ab to tumor cells at acidic pH, the nanoparticle disassembles to CaO 2 and BPQD; CaO 2 produces COMT Ca 2+ and H 2 O 2 , while H 2 O 2 makes oxygen (O 2 ) to promote PDT; under NIR irradiation BPQD facilitates not only the conversion of O 2 to singlet oxygen ( 1 O 2 ) for PDT, but also moderate hyperthermia to accelerate NP dissociation to CaO 2 and BPQD, and conversions of CaO 2 to Ca 2+ and H 2 O 2 , and H 2 O 2 to O 2 , to enhance both COMT and PDT. After supplementary ionomycin treatment to induce intracellular Ca 2+ bursts, the multimodal therapeutics strikingly induce hepatocellular carcinoma apoptosis, likely through the activation of the calpains and caspases 12, 9, and 3, up-regulation of Bax and down-regulation of Bcl-2 proteins. This nanoplatform enables a mutually-amplifying and self-reinforcing synergistic therapy.
研究领域:
癌症治疗
·
纳米技术
·
光动力疗法
·
钙信号
·
药物递送系统
✓ 已验证
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）