ArticleMicrobial cell factories2013
High-yield production of functional soluble single-domain antibodies in the cytoplasm of Escherichia coli.
Article in Microbial cell factories, 2013. The graph could read no effect estimate from its abstract, so it casts no vote on the map. Cited by 36 papers.
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Who cites it
36 citing papers in PubMed.
- Nanobody-based targeted cancer therapy and immunotherapy: fear not the future.Acta pharmacologica Sinica · 2026Review
- Novel recombinant VH43 antibody fragment potentiates the antibacterial activity of cephalosporins against Enterococcus faecium.Archives of microbiology · 2026Article
- An iterative strategy to design 4-1BB agonist nanobodies de novo with generative AI models.Scientific reports · 2025Article
- It's a match: use of the radionuclide theranostic pairEJNMMI radiopharmacy and chemistry · 2025Article
- Scalable, robust, high-throughput expression & purification of nanobodies enabled by 2-stage dynamic control.Metabolic engineering · 2024Article
- Enzyme-Mediated Exponential Glucose Release: A Model-Based Strategy for Continuous Defined Fed-Batch in Small-Scale Cultivations.Bioengineering (Basel, Switzerland) · 2024Article
- Large-Scale Production of Anti-RNase A VHH Expressed inCurrent issues in molecular biology · 2023Article
- Half-Life Extension and Biodistribution Modulation of Biotherapeutics via Red Blood Cell Hitch-Hiking with Novel Anti-Band 3 Single-Domain Antibodies.International journal of molecular sciences · 2022Article
- Review
- The mosaic puzzle of the therapeutic monoclonal antibodies and antibody fragments - A modular transition from full-length immunoglobulins to antibody mimetics.Leukemia research reports · 2022Article
- Targeting ErbB3 Receptor in Cancer with Inhibitory Antibodies from Llama.Biomedicines · 2021Article
- Paper Title "Hu7CG2: A Novel Humanized Anti-Epidermal Growth Factor Receptor (EGFR) Biparatopic Nanobody".Molecular biotechnology · 2021Article
- Nanobodies targeting immune checkpoint molecules for tumor immunotherapy and immunoimaging (Review).International journal of molecular medicine · 2021Review
- Human Hexa-Histidine-Tagged Single-Chain Variable Fragments for Bioimaging of Bacterial Infections.ACS omega · 2021Article
- Application of phage display technology for the production of antibodies against Streptococcus suis serotype 2.PloS one · 2021Article
- Improved production of Humira antibody in the genetically engineered Escherichia coli SHuffle, by co-expression of human PDI-GPx7 fusions.Applied microbiology and biotechnology · 2020Article
- Brain Delivery of Single-Domain Antibodies: A Focus on VHH and VNAR.Pharmaceutics · 2020Review
- Engineering Bacillus subtilis as a Versatile and Stable Platform for Production of Nanobodies.Applied and environmental microbiology · 2020Article
- Synthesis and Characterization of an Epidermal Growth Factor Receptor-Selective RuChembiochem : a European journal of chemical biology · 2020Article
- Versatile Bispidine-Based Bifunctional Chelators forChemistry (Weinheim an der Bergstrasse, Germany) · 2020Article
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Authors and funding
5 authors.
Funding
No grant is acknowledged in the PubMed record.
Abstract
backgroundFor their application in the area of diagnosis and therapy, single-domain antibodies (sdAbs) offer multiple advantages over conventional antibodies and fragments thereof in terms of size, stability, solubility, immunogenicity, production costs as well as tumor uptake and blood clearance. Thus, sdAbs have been identified as valuable next-generation targeting moieties for molecular imaging and drug delivery in the past years. Since these probes are much less complex than conventional antibody fragments, bacterial expression represents a facile method in order to produce sdAbs in large amounts as soluble and functional proteins.
resultsBy the combined use of high cell density cultivation media with a genetically engineered E. coli mutant strain designed for the cytoplasmic formation of proper disulfide bonds, we achieved high level of intracellular sdAb production (up to 200 mg/L). Due to a carboxyterminal hexahistidine epitope, the soluble recombinant sdAbs could be purified by one-step immobilized metal affinity chromatography to apparent homogeneity and easily radiolabeled with 99mTc within 1 h. The intradomain disulfide bridge being critical for the stability and functionality of the sdAb molecule was shown to be properly formed in ~96% of the purified proteins. In vitro binding studies confirmed the high affinity and specificity of the expressed sdAb 7C12 towards its molecular target.
conclusionsOur study demonstrates an efficient cultivation and expression strategy for the production of substantial amounts of soluble and functional sdAbs, which may be adopted for high-yield production of other more complex proteins with multiple disulfides as well.
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