ArticleInternational journal of molecular sciences2014
Refolded scFv antibody fragment against myoglobin shows rapid reaction kinetics.
Article in International journal of molecular sciences, 2014. The graph could read no effect estimate from its abstract, so it casts no vote on the map. Cited by 9 papers.
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Who cites it
9 citing papers in PubMed, 21 citations in OpenAlex.
- Recombinant single-chain variable fragment-based immunoassay for the authentication of Vietnamese ginseng via quantitative detection of majonoside R2.Journal of ginseng research · 2026Article
- Expression and functional characterization of an anti-CD22 scFv targeting B-cell malignancies.Research in pharmaceutical sciences · 2025Article
- Identification of an Immunoglobulin Paratope Binding to Keratan Sulfate and Expression of a Single-Chain Derivative for Imaging.Biomolecules · 2025Article
- Therapeutic potential of a novel IP-10-(anti-HER2 scFv) fusion protein for the treatment of HER2-positive breast cancer.Biotechnology letters · 2023Article
- THETA system allows one-step isolation of tagged proteins through temperature-dependent protein-peptide interaction.Communications biology · 2019Article
- The production of a recombinant tandem single chain fragment variable capable of binding prolamins triggering celiac disease.BMC biotechnology · 2018Article
- Computational de novo design of antibodies binding to a peptide with high affinity.Biotechnology and bioengineering · 2017Article
- Production of an anti-Aβ antibody fragment in Pichia pastoris and in vitro and in vivo validation of its therapeutic effect.PloS one · 2017Article
- Single Chain Variable Fragments Produced in Escherichia coli against Heat-Labile and Heat-Stable Toxins from Enterotoxigenic E. coli.PloS one · 2015Article
Corrections and comments
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Authors and funding
8 authors at 2 institutions in 1 country.
Funding
No grant is acknowledged in the PubMed record.
Abstract
Myoglobin is one of the early biomarkers for acute myocardial infarction. Recently, we have screened an antibody with unique rapid reaction kinetics toward human myoglobin antigen. Antibodies with rapid reaction kinetics are thought to be an early IgG form produced during early stage of in vivo immunization. We produced a recombinant scFv fragment for the premature antibody from Escherichia coli using refolding technology. The scFv gene was constructed by connection of the V(H)-V(L) sequence with a (Gly4Ser)3 linker. The scFv fragment without the pelB leader sequence was expressed at a high level, but the solubility was extremely low. A high concentration of 8 M urea was used for denaturation. The dilution refolding process in the presence of arginine and the redox reagents GSH and GSSH successfully produced a soluble scFv protein. The resultant refolded scFv protein showed association and dissociation values of 9.32 × 10⁻⁴ M⁻¹·s⁻¹ and 6.29 × 10⁻³ s⁻¹, respectively, with an affinity value exceeding 10⁷ M⁻¹ (k(on)/k(off)), maintaining the original rapid reaction kinetics of the premature antibody. The refolded scFv could provide a platform for protein engineering for the clinical application for diagnosis of heart disease and the development of a continuous biosensor.
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