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Radiochemistry in a flash: photochemical conjugation and one-pot radiolabelling of antibodies for immuno-PET
摘要： Monoclonal antibodies (mAbs), immunoglobulin fragments and other proteins are important scaffolds in the development of radiopharmaceuticals for diagnostic immuno-positron emission tomography (immuno-PET) and targeted radioimmunotherapy (RIT). Conventional methods for radiolabelling proteins with metal ions like 68Ga, 64Cu, 89Zr, and 90Y etc require multi-step procedures involving pre-purification, functionalisation with a chelate, and subsequent radiolabelling. Standard coupling chemistries are time consuming, difficult to automate, and involve synthesis, isolation and storage of an intermediate, new molecular entity (the conjugated mAb) whose biochemical properties can differ from those of the parent protein. To circumvent these issues, we developed a photoradiochemical approach that uses fast, chemoselective, light-induced protein modification under mild conditions with novel metal ion binding chelates derivatised with arylazide (ArN3) groups. Experiments show that one-pot photochemical conjugation and radiolabelling of formulated mAbs can be achieved in <20 min.
关键词： Radiochemistry,photochemistry,antibody conjugates,positron emission tomography,arylazides
更新于2025-09-10 09:29:36
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Rapid immunological detection of copper ions using fluorescence immunochromatographic assay
摘要： One-step fluorescence immunochromatographic assay (FICA) has been developed using fluorescent microspheres-labelled monoclonal antibody (McAb) probe for the rapid detection of copper ions in herbal plants and soil samples were investigated in this study. The results can be read by the fluorescence reader according to the fitted standard curve equation. Under optimal conditions, detecting copper ions with FICA can be done within 15 min with a LOD (limit of detection) of 0.2 μg/L, and was very useful for the rapid onsite testing. A total of 31 solid samples were pre-treated by microwave digestion combined with cation exchange solid phase extraction and prepared as test samples for FICA and ICP-AES. The two methods showed good relativity with correlation coefficients of 0.924 and 0.969 in herbal plants and soil samples for copper detection, respectively. We hope that our research can help promote the daily monitoring of heavy metal pollution effectively.
关键词： Copper ions,fluorescence immunochromatographic assay,monoclonal antibody,rapid onsite testing
更新于2025-09-09 09:28:46
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Rapid immunofluorescence assay for staphylococcal enterotoxin A using magnetic nanoparticles
摘要： A competitive immunoassay for staphylococcal enterotoxin A (SEA) detection in milk was developed, using immobilised antibody onto magnetic nanoparticles (MNPs). MNPs were prepared and then modified to introduce amino groups on them. The morphology and size of the obtained both unmodified and modified MNPs were characterized using TEM analyses. Monoclonal anti-SEA antibody was immobilised onto the modified MNPs (MNP-Ab). Staphylococcal enterotoxin A was conjugated with fluorescent dye ATTO620NHS. The characteristics of fluorescence conjugate were examined. The amount of MNP-Ab and concentration of the fluorescent conjugate used for competitive immunoassay were optimized: 0.25 mg and 53 lg mL(cid:1)1, respectively. The detection limit of developed immunoassay was determined – 0.23 ng mL(cid:1)1 SEA in spiked milk samples. The immunoassay takes only 30 min, the magnetic separation is fast (<10 s) and the volume of the sample for analysis is very small (200 lL).
关键词： fluorescent conjugate,staphylococcal enterotoxin,milk,Antibody immobilisation,magnetic nanoparticles,immunofluorescence
更新于2025-09-09 09:28:46
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Activatable Near-Infrared Fluorescence Imaging Using PEGylated Bacteriochlorin-Based Chlorin and BODIPY-Dyads as Probes for Detecting Cancer
摘要： Near infrared (NIR) fluorescent probes are attractive tools for biomedical in vivo imaging due to the relatively deeper tissue penetration and lower background autofluorescence. Activatable probes are turned on only after binding to their target, further improving target to background ratios. However, the number of available activatable NIR probes is limited. In this study, we introduce two types of activatable NIR fluorophores derived from bacteriochlorin; chlorin-bacteriochlorin energy-transfer dyads and boron-dipyrromethene (BODIPY)-bacteriochlorin energy-transfer dyads. These fluorophores are characterized by multiple narrow excitation bands with relatively strong emission in the NIR. Targeted bacteriochlorin-based antibody or peptide probes have been previously limited by aggregation after conjugation. Polyethylene glycol (PEG) chains were added to improve the hydrophilicity without altering pharmacokinetics of the targeting moieties. These PEGylated bacteriochlorin-based activatable fluorophores have potential as targeted activatable, multi-color NIR fluorescent probes for in vivo applications.
关键词： Monoclonal antibody,Molecular imaging,Bacteriochlorin,Cancer
更新于2025-09-09 09:28:46
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Regulation of osteoblast differentiation by osteocytes cultured on sclerostin antibody conjugated TiO2 nanotube array
摘要： Sclerostin is a negative regulator of the Wnt signaling pathway for osteoblast differentiation. In this study, osteoblasts were co-cultured with osteocytes (MLO-Y4 cells) on the surface of sclerostin antibody-conjugated TiO2 nanotube arrays (TNTs-scl). Field emission scanning electron microscopy (SEM), contact angle measurement and confocal laser scanning microscope (CLSM) were employed to characterize the conjugation of sclerostin antibody onto the surface of TiO2 nanotube arrays. The cellular viability and morphology results displayed TNTs-scl (TNT30-scl and TNT70-scl) were beneficial to the growth of MLO-Y4 cells. There was no apparent change in sclerostin gene expression between MLO-Y4 cells grown on TNTs and TNTs-scl. However, TNTs-scl significantly reduced the amount of sclerostin in the medium. In comparison with the control groups, osteoblasts displayed higher differentiation capability when co-cultured with MLO-Y4 cells on the surface TNTs-scl, which was indicated by the ALP activity, mineralization capability as well as expression levels of key proteins in Wnt signaling. This study provides a simple strategy to engineer titanium surface for bone fracture recovery, especially in osteoporotic conditions.
关键词： TiO2 nanotube arrays,osteocytes,osteogenic differentiation,Sclerostin antibody
更新于2025-09-04 15:30:14
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Multiplex body fluid identification using surface plasmon resonance imaging with principal component analysis
摘要： Body ?uid identi?cation is a key component in forensic casework, providing important information for the reconstruction of criminal events. Body ?uid identi?cation in combination with DNA analysis allows the linking of individuals to criminal acts and can therefore be of great importance in determining the outcome of criminal court cases. However, none of the current body ?uid identi?cation methods meet all forensic requirements, such as a high sensitivity, a high speci?city and the ability to analyse multiple body ?uids in a single run. In this pilot-study, we explore, for the ?rst time, surface plasmon resonance imaging (SPRi) with antibody-based detection to serve as a novel multiplex body ?uid identi?cation assay for blood, semen, saliva, urine and sweat using minimal sample preparation. A training set consisting of ten donors per body ?uid was analysed to determine whether body ?uid speci?c response signals could be obtained. Principal component analysis (PCA) was performed as a statistical tool to cluster the body ?uid samples by response signal pattern reduction and to uncover the sources of variation between the body ?uids. Four principal components allowed complete clustering of all body ?uid types. Blind testing of body ?uid samples revealed that ?ve out of eight unknown samples could correctly be clustered to their corresponding group, three out of eight samples were identi?ed as inconclusive. Although optimization of the current SPRi method is required for use in the forensic ?eld, this pilot-study demonstrates the feasibility of SPRi to di?erentiate ?ve forensically relevant body ?uids.
关键词： Body ?uid identi?cation,Antibody,Forensic science,Surface plasmon resonance (SPR),Principal component analysis (PCA)
更新于2025-09-04 15:30:14