ArticleJournal of translational medicine2014
Construction of a synthetic phage-displayed Nanobody library with CDR3 regions randomized by trinucleotide cassettes for diagnostic applications.
Article in Journal of translational medicine, 2014. The graph could read no effect estimate from its abstract, so it casts no vote on the map. Cited by 58 papers.
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Who cites it
58 citing papers in PubMed, 112 citations in OpenAlex.
- Review
- Article
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- Nanobodies: From Discovery to AI-Driven Design.Biology · 2025Review
- Validation and Optimization of PURE Ribosome Display for Screening Synthetic Nanobody Libraries.Antibodies (Basel, Switzerland) · 2025Article
- A synthetic heavy chain variable domain antibody library (VHL) provides highly functional antibodies with favorable developability.Protein science : a publication of the Protein Society · 2025Article
- On the origin of non-specific binders isolated in the selection of phage display peptide libraries.Frontiers in microbiology · 2025Review
- A new strategy to generate nanobodies for the coumaphos based on the synthesized nanobody libraries.Food chemistry · 2024Article
- The process using a synthetic library that generates multiple diverse human single domain antibodies.Antibody therapeutics · 2024Article
- Single-domain antibodies against SARS-CoV-2 RBD from a two-stage phage screening of universal and focused synthetic libraries.BMC infectious diseases · 2024Article
- Development of a synthetic library of humanized nanobodies for targeted IL-6 inhibition.Frontiers in bioengineering and biotechnology · 2024Article
- Screening and construction of nanobodies against human CD93 using phage libraries and study of their antiangiogenic effects.Frontiers in bioengineering and biotechnology · 2024Article
- Lymphocyte-activation gene 3 in cancer immunotherapy: function, prognostic biomarker and therapeutic potentials.Frontiers in immunology · 2024Review
- Shark IgNAR: The Next Broad Application Antibody in Clinical Diagnoses and Tumor Therapies?Marine drugs · 2023Review
- Ultrasensitive Electrochemical Immunosensors Using Nanobodies as Biocompatible Sniffer Tools of Agricultural Contaminants and Human Disease Biomarkers.Micromachines · 2023Review
- Design of Oscillatory Networks through Post-Translational Control of Network Components.Synthetic biology and engineering · 2023Article
- Design and Construction of a Synthetic Nanobody Library: Testing Its Potential with a Single Selection Round Strategy.Molecules (Basel, Switzerland) · 2023Article
- Bacteriophages as Biotechnological Tools.Viruses · 2023Review
- Construction of Synthetic VHH Libraries in Ribosome Display Format.Methods in molecular biology (Clifton, N.J.) · 2023Article
- Construction of Synthetic Antibody Phage Display Libraries.Methods in molecular biology (Clifton, N.J.) · 2023Article
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Authors and funding
5 authors at 2 institutions in 1 country.
Funding
No grant is acknowledged in the PubMed record.
Abstract
backgroundNanobodies (Nbs) have proved their great value as therapeutic molecules and clinical diagnostic tools. Although the routine procedure to obtain Nbs is to immunize camels with antigens, it is unavailable to immunize a camel when the antigens are highly toxic, pathogenic or nonimmunogenic. A synthetic phage display library is an alternative to generate Nbs against such targets, besides all the other ones.
methodsWe constructed a large and diverse synthetic phage display Nanobody (Nb) library based on the conserved camel single-domain antibody fragment (VHH) framework of cAbBCII10. Diversity was introduced in the complementarity-determining region 3 (CDR3) by means of randomization of synthetic oligonucleotides. Then human prealbumin (PA) and neutrophil gelatinase-associated lipocalin (NGAL) were used to select specific Nbs from this library. Furthermore, a sandwich enzyme-linked immunosorbent assay (ELISA) was developed to detect PA based on horseradish peroxidase (HRP)-conjugated anti-PA Nb isolated from this study and another biotinylated anti-PA Nb obtained from an immune library, in our previous study.
resultsA large and diverse synthetic phage display Nb library with CDR3 regions randomized by trinucleotide cassettes was constructed. The library size was 1.65 × 10(9) CFU/mL and the correct insertion ratio was nearly 100%. A Nb against human PA and against NGAL was successfully isolated from the synthetic library. The obtained anti-PA Nb was effectively used to develop a sandwich ELISA for PA detection and it demonstrated a working range from 50 to 1000 ng/mL, with a limit of detection (LOD) of 27.1 ng/mL.
conclusionThis proposed novel synthetic library was a good source for obtaining some antigen-specific Nbs. This approach could provide crucial support to an immune library and a naïve library in the acquisition of specific Nbs, potentially functioning as a great resource for medical diagnostic applications. In addition, we have successfully developed a novel sandwich ELISA to detect PA, which could provide great assistance for clinical PA detection.
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