ArticleMicrobial cell factories2014
Bacterial cytoplasm as an effective cell compartment for producing functional VHH-based affinity reagents and Camelidae IgG-like recombinant antibodies.
Article in Microbial cell factories, 2014. The graph could read no effect estimate from its abstract, so it casts no vote on the map. Cited by 29 papers, 1 of them a synthesis that pooled it.
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Who cites it
29 citing papers in PubMed, 1 synthesis or guideline pooled it, 73 citations in OpenAlex.
- Research Progress and Applications of Multivalent, Multispecific and Modified Nanobodies for Disease Treatment.Frontiers in immunology · 2021Pooled it
- Parallelised Cloning, Mammalian Cell Expression, and Purification of Nanobodies Identified by Phage Display.Bio-protocol · 2026Article
- Production of Antibody Fragments Suitable for 1:1, Site-Specific Labeling.Methods in molecular biology (Clifton, N.J.) · 2026Article
- Recent advances in recombinant production of soluble proteins in E. coli.Microbial cell factories · 2025Review
- Urinary Extracellular Vesicles as a Readily Available Biomarker Source: A Simplified Stratification Method.International journal of molecular sciences · 2024Article
- A Practical Guide for the Quality Evaluation of Fluobodies/Chromobodies.Biomolecules · 2024Article
- Novel bispecific nanobody mitigates experimental intestinal inflammation in mice by targeting TNF-α and IL-23p19 bioactivities.Clinical and translational medicine · 2024Article
- Review
- Biological Applications of Synthetic Binders Isolated from a Conceptually New Adhiron Library.Biomolecules · 2023Article
- Native Agarose Gels and Contact Blotting as Means to Optimize the Protocols for the Formation of Antigen-Ligand Complexes.Bioengineering (Basel, Switzerland) · 2023Article
- DNA-Directed Protein Anchoring on Oligo/Alkanethiol-Coated Gold Nanoparticles: A Versatile Platform for Biosensing Applications.Nanomaterials (Basel, Switzerland) · 2022Article
- Research progress and applications of nanobody in human infectious diseases.Frontiers in pharmacology · 2022Review
- Native llama Nanobody Library Panning Performed by Phage and Yeast Display Provides Binders Suitable for C-Reactive Protein Detection.Biosensors · 2021Article
- CDR1 Composition Can Affect Nanobody Recombinant Expression Yields.Biomolecules · 2021Article
- Recent Advancements in Receptor Layer Engineering for Applications in SPR-Based Immunodiagnostics.Sensors (Basel, Switzerland) · 2021Review
- Effect of Humanizing Mutations on the Stability of the Llama Single-Domain Variable Region.Biomolecules · 2021Article
- Advances in mesenchymal stem cell exosomes: a review.Stem cell research & therapy · 2021Review
- Optimization of Methods for the Production and Refolding of Biologically Active Disulfide Bond-Rich Antibody Fragments in Microbial Hosts.Antibodies (Basel, Switzerland) · 2020Article
- Recombinant expression of nanobodies and nanobody-derived immunoreagents.Protein expression and purification · 2020Review
- The novel llama-human chimeric antibody has potent effect in lowering LDL-c levels in hPCSK9 transgenic rats.Clinical and translational medicine · 2020Article
Corrections and comments
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Authors and funding
9 authors at 4 institutions in 3 countries.
Funding
No grant is acknowledged in the PubMed record.
Abstract
backgroundThe isolation of recombinant antibody fragments from displayed libraries represents a powerful alternative to the generation of IgGs using hybridoma technology. The selected antibody fragments can then be easily engineered into (multi)-tagged constructs of variable mass and complexity as well as reconstituted into Camelidae IgG-like molecules when expressed fused to Fc domains. Nevertheless, all antibody constructs depend on an oxidizing environment for correct folding and consequently still belong to the proteins difficult to express in bacteria. In such organisms they are mostly produced at low yields in the periplasmic space.
resultsWe demonstrate that fusion constructs of recombinant antibodies in combination with multiple tags can be produced at high yields and totally functional in the cytoplasm of bacteria expressing sulfhydryl oxidase. The method was applied to structurally demanding molecules such as VHHs fused to SNAP and Fc domains and was validated using the antibody-derived reagents in a variety of immune techniques (FACS, ELISA, WB, IP, SPR, and IF).
conclusionsThe collected data demonstrate the feasibility of a method that establishes a totally new approach for producing rapidly and inexpensively functional Camelidae IgG-like monoclonal antibodies and antibody-based reagents containing multiple disulfide bonds and suitable for both basic research and clinical applications.
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Registered trials
Read under generation 80e0d062 · epoch 390. Bibliography from PubMed, PubMed Central and OpenAlex; grants from NIH RePORTER; trial links from ClinicalTrials.gov; estimates, votes and beliefs from the OpenQuestion graph.