Mesothelin, a cancer biomarker overexpressed in tumors of epithelial origin, is a target for nanotechnology-based diagnostic, therapeutic, and prognostic applications. The currently available anti-mesothelin antibodies present limitations, including low penetration due to large size and/or lack of in vivo stability. Single domain antibodies (sdAbs) or nanobodies (Nbs) provide powerful solutions to these specific problems. We generated a phage-display library of Nbs that were amplified from B cells of a llama that was immunized with human recombinant mesothelin. Two nanobodies (Nb A1 and Nb C6) were selected on the basis of affinity (K(D) = 15 and 30 nM, respectively). Nb A1 was further modified by adding either a cysteine to permit maleimide-based bioconjugations or a sequence for the site-specific metabolic addition of a biotin in vivo. Both systems of conjugation (thiol-maleimide and streptavidin/biotin) were used to characterize and validate Nb A1 and to functionalize nanoparticles. We showed that anti-mesothelin Nb A1 could detect native and denatured mesothelin in various diagnostic applications, including flow cytometry, western blotting, immunofluorescence, and optical imaging. In conclusion, anti-mesothelin Nbs are novel, cost-effective, small, and single domain reagents with high affinity and specificity for the tumor-associated antigen mesothelin, which can be simply bioengineered for attachment to nanoparticles or modified surfaces using multiple bioconjugation strategies. These anti-mesothelin Nbs can be useful in both conventional and nanotechnology-based diagnostic, therapeutic and prognostic biomedical applications.
Download full-text PDF |
Source |
|---|---|
| http://dx.doi.org/10.1166/jbn.2015.2063 | DOI Listing |
Publication Analysis
Top Keywords
Similar Publications
Unsupervised self-organising map classification of Raman spectra from prostate cell lines uncovers substratified prostate cancer disease states.
Sci Rep
January 2025
School of Physics, Engineering and Technology, University of York, Heslington, York, YO10 5DD, UK.
Prostate cancer is a disease which poses an interesting clinical question: Should it be treated? Only a small subset of prostate cancers are aggressive and require removal and treatment to prevent metastatic spread. However, conventional diagnostics remain challenged to risk-stratify such patients; hence, new methods of approach to biomolecularly sub-classify the disease are needed. Here we use an unsupervised self-organising map approach to analyse live-cell Raman spectroscopy data obtained from prostate cell-lines; our aim is to exemplify this method to sub-stratify, at the single-cell-level, the cancer disease state using high-dimensional datasets with minimal preprocessing.
View Article and Find Full Text PDFOpen challenges and opportunities in federated foundation models towards biomedical healthcare.
BioData Min
January 2025
School of Computer Science, Fudan University, Shanghai, China.
This survey explores the transformative impact of foundation models (FMs) in artificial intelligence, focusing on their integration with federated learning (FL) in biomedical research. Foundation models such as ChatGPT, LLaMa, and CLIP, which are trained on vast datasets through methods including unsupervised pretraining, self-supervised learning, instructed fine-tuning, and reinforcement learning from human feedback, represent significant advancements in machine learning. These models, with their ability to generate coherent text and realistic images, are crucial for biomedical applications that require processing diverse data forms such as clinical reports, diagnostic images, and multimodal patient interactions.
View Article and Find Full Text PDFBacterial proteome microarray technology in biomedical research.
Trends Biotechnol
January 2025
Department of Food Safety/Hygiene and Risk Management, National Cheng Kung University, Tainan, Taiwan; Institute of Basic Medical Sciences, National Cheng Kung University, Tainan, Taiwan. Electronic address:
Bacterial proteome microarrays are high-throughput, adaptable tools that allow the simultaneous investigation of thousands of proteins from various bacterial species. These arrays are used to explore bacterial pathogenicity, pathogen-host interactions, and clinical diseases. Recent advancements have expanded their application to profiling human antibodies, identifying biomarkers for infectious and autoimmune diseases, and studying antimicrobial peptides (AMPs).
View Article and Find Full Text PDFSynthesis of BODIPYs using organoindium reagents and survey of their cytotoxicity and cell uptake on nervous system cells.
Bioorg Chem
December 2024
Universidade da Coruña, CICA - Centro Interdisciplinar de Química e Bioloxía and Departamento de Química, Facultad de Ciencias, Campus A Zapateira, 15071 A Coruña, Spain. Electronic address:
In this study, a series of BODIPY dyes were synthesized, containing various substituents at meso position. Further functionalization of the BODIPY framework at C2 and C2-C6 position(s) by palladium-catalysed cross-coupling reactions using organoindium reagents (RIn) was efficiently assessed, starting from C2(6)-halogenated BODIPYs, and their optical properties were measured. The cytotoxicity of BODIPY dyes on SH-SY5Y neuronal cells by MTT assay showed that those compounds bearing thien-2-yl and benzonitrile moieties at meso position, exhibited great efficiency in maintaining cell viability under all tested conditions (up to 50 µM for 24 h and 48 h).
View Article and Find Full Text PDFThe Impact of Pistacia khinjuk plant gender on silver nanoparticle synthesis: Are extracts of root obtained from female plants preferentially used?
Biochem Biophys Res Commun
December 2024
Department of Nanotechnology, Institute of Science, Dicle University, 21280, Diyarbakir, Turkey. Electronic address:
Pistacia khinjuk, a dioecious plant native to Southeast Anatolia, Turkey, features distinct male and female individuals with varying bioactive compound profiles. This study investigates the gender-specific phytochemical composition of root extracts from male and female Pistacia khinjuk plants and their influence on the green synthesis of silver nanoparticles. Using natural bioactive compounds such as polyphenols, flavonoids, alkaloids, and terpenoids as reducing and stabilizing agents, the study demonstrates significant differences between the nanoparticles synthesized from male and female root extracts.
View Article and Find Full Text PDFWant AI Summaries of new PubMed Abstracts delivered to your In-box?
Enter search terms and have AI summaries delivered each week - change queries or unsubscribe any time!