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Dual-Modality ImmunoPET/Fluorescence Imaging of Prostate Cancer with an Anti-PSCA Cys-Minibody
摘要: Inadequate diagnostic methods for prostate cancer lead to over- and undertreatment, and the inability to intraoperatively visualize positive margins may limit the success of surgical resection. Prostate cancer visualization could be improved by combining the complementary modalities of immuno-positron emission tomography (immunoPET) for preoperative disease detection, and fluorescence imaging-guided surgery (FIGS) for real-time intraoperative tumor margin identification. Here, we report on the evaluation of dual-labeled humanized anti-prostate stem cell antigen (PSCA) cys-minibody (A11 cMb) for immunoPET/fluorescence imaging in subcutaneous and orthotopic prostate cancer models. Methods: A11 cMb was site-specifically conjugated with the near-infrared fluorophore Cy5.5 and radiolabeled with 124I or 89Zr. 124I-A11 cMb-Cy5.5 was used for successive immunoPET/fluorescence imaging of prostate cancer xenografts expressing high or moderate levels of PSCA (22Rv1-PSCA and PC3-PSCA). 89Zr-A11 cMb-Cy5.5 dual-modality imaging was evaluated in an orthotopic model. Ex vivo biodistribution at 24 h was used to confirm the uptake values, and tumors were visualized by post-mortem fluorescence imaging. Results: A11 cMb-Cy5.5 retained low nanomolar affinity for PSCA-positive cells. Conjugation conditions were established (dye-to-protein ratio of 0.7:1) that did not affect the biodistribution, pharmacokinetics, or clearance of A11 cMb. ImmunoPET using dual-labeled 124I-A11 cMb-Cy5.5 showed specific targeting to both 22Rv1-PSCA and PC3-PSCA s.c. xenografts in nude mice. Ex vivo biodistribution confirmed specific uptake to PSCA-expressing tumors with 22Rv1-PSCA:22Rv1 and PC3-PSCA:PC3 ratios of 13:1 and 5.6:1, respectively. Consistent with the immunoPET, fluorescence imaging showed a strong signal from both 22Rv1-PSCA and PC3-PSCA tumors compared with non-PSCA expressing tumors. In an orthotopic model, 89Zr-A11 cMb-Cy5.5 immunoPET was able to detect intraprostatically implanted 22Rv1-PSCA cells. Importantly, fluorescence imaging clearly distinguished the prostate tumor from surrounding seminal vesicles. Conclusion: Dual-labeled A11 cMb specifically visualized PSCA-positive tumor by successive immunoPET/fluorescence, which can potentially be translated for preoperative whole-body prostate cancer detection and intraoperative surgical guidance in patients.
关键词: molecular imaging,immunoPET,prostate cancer,antibody fragment,fluorescence
更新于2025-09-23 15:22:29
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Photoluminescence Immunosensor Based on Bovine Leukemia Virus Proteins Immobilized on the ZnO Nanorods
摘要: Bovine leukaemia virus (BLV) proteins gp51, which are serving as antigens for specific antibodies against BLV proteins (anti-gp51), were applied as biological recognition part in the design of immunosensor devoted for the determination of anti-gp51. The efficiency of the immobilization of BLV proteins gp51 on ZnO nanorod (ZnO-NR) modified glass (ZnO-NR/glass) surface was evaluated. The formation of antigen-antibody complex on the ZnO/glass modified by the BLV proteins gp51 (gp51/ZnO-NR/glass) was investigated by the determination of changes in ZnO photoluminescence. The applicability of gp51/ZnO-NR/glass in the design of photoluminescence based immunosensor was evaluated. Bovine serum albumin (BSA) was applied for the modification of sensing gp51 layer in order to form gp51&BSA layer with advanced selectivity. Polyallylamine hydrochloride (PAH) was applied in order to improve the immobilization of gp51 and BSA based sensing layer (gp51&BSA) on the surface of ZnO-NR/glass. PAH was applied during the formation of gp51&BSA/PAH/ZnO-NR/glass structure. Some aspects of the mechanism of interaction between biomolecules (gp51, bovine serum albumin (BSA) and anti-gp51) and ZnO-NR during the preparation and action of gp51&BSA/ZnO-NR/glass- and gp51&BSA/PAH/ZnO-NR/glass-based immunosensors have been discussed.
关键词: Bovine leukemia virus (BLV),Photoluminescence,Optical immunosensor,ZnO nanorods,Antigen-antibody complex.
更新于2025-09-23 15:22:29
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Photoimmunoconjugates: novel synthetic strategies to target and treat cancer by photodynamic therapy
摘要: Photodynamic therapy (PDT) combines a photosensitizer (PS) with the physical energy of non-ionizing light to trigger cell death pathways. PDT has potential as a therapeutic modality to be used in alternative or in combination with other conventional cancer treatment protocols (e.g. surgery, chemotherapy and radiotherapy). Still, due to the lack of specificity of the current PSs to target the tumor cells, several studies have exploited their conjugation with targeting moieties. PSs conjugated with antibodies (Abs) or their fragments, able to bind antigens overexpressed in the tumors, have demonstrated potential in PDT of tumors. This review provides an overview of the most recent advances on photoimmunoconjugates (PICs) for cancer PDT, which involve the first and second-generation PSs conjugated to Abs. This is an update of our previous review “Antibodies armed with photosensitizers: from chemical synthesis to photobiological applications”, published in 2015 in Org. Biomol. Chem.
关键词: Photoimmunoconjugates,Antibody conjugation,Photosensitizers,Cancer treatment,Photodynamic therapy
更新于2025-09-23 15:22:29
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Site-specific chelator-antibody conjugation for PET and SPECT imaging with radiometals
摘要: Antibodies and their derivatives radiolabelled with positron- and gamma-emitting radiometals enable sensitive and quantitative molecular Positron Emission Tomography (PET) and Single Photon Emission Computed Tomography (SPECT) imaging of antibody distribution in vivo. Chelators that are covalently attached to antibodies allow radiolabelling with metallic PET and SPECT radioisotopes. Conventional strategies for chelator-protein conjugation generate heterogeneous mixtures of bioconjugates that can exhibit reduced affinity for their targets, and undesirable biodistribution and pharmacokinetics. Recent advances in bioconjugation technology enable site-specific modification to generate well-defined constructs with superior properties. Herein we survey existing site-specific chelator-protein conjugation methods. These include chelator attachment to cysteines/disulfide bonds or the glycan region of the antibody, enzyme-mediated chelator conjugation, and incorporation of sequences of amino acids that chelate the radiometal. Such technology will allow better use of PET and SPECT imaging in the development of antibody-based therapies.
关键词: antibody,radiometals,PET,chelator,site-specific conjugation,SPECT,molecular imaging
更新于2025-09-23 15:21:21
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[Environmental Chemistry for a Sustainable World] Nanosensors for Environmental Applications Volume 43 || Development of Optical Sensor Strips for Point-of-Care Testing for Pesticide
摘要: Disposable or point-of-care sensors are a promising tool for low-cost and rapid sensing of analytes including pesticides. They find important applications in pesticide-contaminated food, agro-products, and water quality monitoring. This chapter highlights the implication and significance of pesticide residue identification in foodstuffs and overviews the most frequently engaged analytical techniques, and finally their benefits and limitations are discussed. Disposable strip-based biosensors have their intrinsic advantages and some disadvantages, but their cost-effectiveness and portability have turned them as a potential possibility for point-of-care (POC) testing of various pesticides. The fabrication of robust, low-cost, reliable, and sensitive sensors with the aid of both simple naked eye-based and portable readout-based detectors is the driving factor in this sensor’s technology area. The pending limitations can be overcome by adapting new specific recognition elements and better signal generative particles or systems. The integration of these devices with card readers or smartphones can make them more user-friendly and will provide more accurate quantitative information.
关键词: Organophosphates,Pesticide,Immunoassay,Aptamer,Biosensors,Immunochromatographic assay,Point-of-care,Antibody,Rapid detection,Nanosensors,Gas chromatography
更新于2025-09-23 15:21:01
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Time-resolved fluorescent immunochromatographic assay-based on three antibody labels for the simultaneous detection of aflatoxin B <sub/>1</sub> and zearalenone in Chinese herbal medicines
摘要: A time-resolved fluorescent immunochromatographic assay (TRFICA) was successfully developed for the sensitive, simultaneous, and quantitative detection of aflatoxin B1 (AFB1) and zearalenone (ZEN) in Chinese herbal medicines. Eu-nanospheres (EuNPs) with unique optical properties increased the stability and sensitivity of the immunochromatographic assay. To obtain stable quantitative results, we applied a three-label system in which monoclonal antibodies for AFB1 and ZEN were conjugated to the EuNPs as detection probes on the test line (T line), and EuNP-labelled chicken IgY conjugates acted as the reference on the control line (C line). The fluorescence intensities of the T and C lines were recorded, and the T/C ratio was employed as the quantitative signal for the elimination of strip variation and matrix effects. The parameters that affected the TRFICA were optimised. Under optimal conditions, the established TRFICA gave good linear ranges from 0.60 μg/kg to 3.92 μg/kg for AFB1 and from 0.40 μg/kg to 1.28 μg/kg for ZEN. The limits of detection for AFB1 and ZEN were as low as 0.60 and 0.40 μg/kg, respectively, in Chinese herbal medicines Semen coicis, Rhizoma dioscoreae, and Platycodon grandiflorus, respectively. The average recoveries of the spiked samples were 73%–95% for AFB1 and 75.83%–90% for ZEN, both with a relative standard deviation of < 9.08%. The results of 15 actual samples detected by the developed TRFICA showed a satisfactory correlation with those of ultra-performance liquid chromatography tandem mass spectrometry. Therefore, the TRFICA is a simple, rapid, and sensitive approach to quantitatively detect mycotoxins in Chinese herbal medicines.
关键词: aflatoxin B1,time-resolved fluorescent immunochromatographic assay,zearalenone,Three antibody labels,Chinese herbal medicines
更新于2025-09-23 15:21:01
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Screening and characterisation of CdTe/CdS quantum dot-binding peptides for material surface functionalisation
摘要: Quantum dots (QDs) are promising nanomaterials due to their unique photophysical properties. For them to be useful in biological applications, the particle surface generally needs to be conjugated to biological molecules, such as antibodies. In this study, we screened CdTe/CdS QD-binding peptides from a phage display library as linkers for simple and bio-friendly QD modification. Among five QD-binding peptide candidates, a series of truncated peptides designed from two high-affinity peptides were subjected to an array-based binding assay with QDs to assess their functional core sequences and characteristics. Linking these isolated, shortened peptides (PWSLNR and SGVYK) with an antibody-binding peptide (NKFRGKYK) created dual-functional peptides that are capable of QD surface functionalisation by antibodies. Consequently, the dual-functional peptides could mediate anti-CD9 antibody functionalisation onto CdTe/CdS QD surface; CD9 protein imaging of cancer cells was also demonstrated. Our proposed peptides offer an effective vehicle for QD surface functionalisation in biological applications.
关键词: surface functionalisation,Quantum dots,bioimaging,peptides,CdTe/CdS,phage display,antibody conjugation
更新于2025-09-23 15:19:57
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Fluorometric immunoassay for the simultaneous determination of the tumor markers carcinoembryonic antigen and cytokeratin 19 fragment using two kinds of CdSe/ZnS quantum dot nanobeads and magnetic beads
摘要: A method is described for the simultaneous determination of the carcinoembryonic antigen (CEA) and cytokeratin 19 fragment (CYFRA21-1). Two kinds of CdSe/ZnS quantum dot nanobeads (QBs), with emission maxima at 530 nm (green) and 585 nm (yellow), were used as labels, and magnetic beads (MBs) for separation. The MBs were used as substrates to couple CEA and CYFRA21-1 antibody for isolating the proteins. Then, the differently colored QBs were linked to the antibodies against CEA and CYFRA21-1, respectively. Following the formation of the immunocomplex, the intensities of the green and yellow emissions were measured at the same excitation wavelength of 340 nm. The detection limits are 0.1 ng·mL?1 for CEA, and of 0.2 ng·mL?1 for CYFRA21-1. The recoveries from spiked serum are 92.1 - 118.1% for CEA, and from 90.8% to 115.2% for CYFRA21-1, with the relative standard deviations of 6.3 - 12.3% and 7.1 - 11.8%. The method was successfully applied to the simultaneous determination of the two proteins in human serum sample (n = 45). The results correlated well with those of the chemiluminescent enzyme immunoassay kit.
关键词: Lung cancer,Human serum,Fluorescence,Multiplexed detection,Antibody
更新于2025-09-23 15:19:57
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Sandwich magnetically imprinted immunosensor for electrochemiluminescence ultrasensing diethylstilbestrol based on enhanced luminescence of Ru@SiO2 by CdTe@ZnS quantum dots
摘要: A molecularly imprinted magnetic sensor with electroluminescent tags (MIP-ECL sensor) was developed for ultrasensing diethylstilbestrol (DES). A strategy is exploited to enhance ECL emission of the [Ru(bpy)3]2 +-tripropyl amine (TPrA) system by CdTe@ZnS quantum-dots (QDs) through energy transfer. Magnetically molecularly imprinted nanoparticles (MMIPs NPs) based on Fe3O4@SiO2 carriers are artificial, easily reproducible, and could replace easily inactivated first antibodies for capturing more DES molecules. Functionalized bio-conjugates of single antibody-CdTe@ZnS (Ab-CdTe@ZnS) are for the first time loaded on signal labels of Ru(bpy)3 2 +-doped silica nanocomposites (Ru@SiO2) for signal amplification. The final bio-conjugated signal probes are denoted as Ab-DES/CdTe@ZnS-Ru@SiO2. MMIPs beads that have captured antigens are bio-conjugated with antibody-labeled luminescent probes by specific immunoreactive reaction, and then the luminescent immunocomplex generates ECL signal on the magnetic electrode. The logarithm of ECL intensities depend linearly on the logarithm of DES concentrations in the range from 4.8×10? 4 to 36.0 nM with a detection limit of 0.025 pM. This novel assay is much more sensitive than other MIP sensors, and achieves lower cost and more enhanced stability than other immunosensors. The sensor is significantly potential and has been applied to DES detection in actual environment.
关键词: [Ru(bpy)3]2+-doped silica,Probe-mode ECL sensor,Core-shell QDs,Magnetically imprinted nanoparticles,Diethylstilbestrol,Single antibody sandwich-type
更新于2025-09-23 15:19:57
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Development of Carbon Quantum Dota??Labeled Antibody Fluorescence Immunoassays for the Detection of Morphine in Hot Pot Soup Base
摘要: Morphine is the most representative characteristic alkaloid existed in the extract solution of poppy shell. The analysis of morphine can be used to judge whether poppy shell is illegally added to the hot pot soup base. A sensitive, environmental-friendly carbon quantum dots (C-Dots)–based fluorescence immunoassay (C-Dots-FLISA) was developed to detect morphine (MOP). Novel water-soluble C-Dots were synthesized using a one-step hydrothermal reaction, with citric acid serving as the carbon source and ethylene diamine acting as the nitrogen source. Moreover, the C-Dots displayed blue fluorescence with an emission peak at 440 nm (350 nm excitation), which also showed good stability. C-Dots which contained amine were used to conjugate with anti-morphine antibody by glutaraldehyde as coupling reagents. The anti-morphine antibody–labeled C-Dots (Abs-labeled C-Dots) were characterized by fluorescence spectrum, transmission electron microscopy (TEM), and gel electrophoresis. C-Dots-FLISA was developed and applied to quantitative detection of morphine. Under the optimal conditions, the linear range spanned from 3.2 × 10?4 to 10 mg/L (R2 = 0.992), and the detection limit was 3 × 10?4 mg/L. These results demonstrated that the developed C-Dots-FLISA could be applied as a sensitive and convenient tool for rapid detection of morphine.
关键词: Fluorescence immunoassay,Carbon quantum dots,Anti-morphine antibody,Morphine,Hot pot soup base
更新于2025-09-23 15:19:57