ArticleApplied and environmental microbiology2002
Functional production and characterization of a fibrin-specific single-chain antibody fragment from Bacillus subtilis: effects of molecular chaperones and a wall-bound protease on antibody fragment production.
Article in Applied and environmental microbiology, 2002. The graph could read no effect estimate from its abstract, so it casts no vote on the map. Cited by 75 papers.
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Who cites it
75 citing papers in PubMed, 198 citations in OpenAlex.
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- CRISPR/Cas9-engineered Bacillus subtilis chassis for tailored chitooligosaccharide production from marine waste chitosan.Microbial cell factories · 2025Article
- A novel signal peptide toolbox for optimized heterologous expression of yeast pheromones in Bacillus subtilis.Journal, genetic engineering & biotechnology · 2025Article
- Bacterial two-hybrid systems evolved: innovations for protein-protein interaction research.Journal of bacteriology · 2025Review
- Expression and Biological Activity Analysis of Recombinant Fibronectin3 Protein inBiotech (Basel (Switzerland)) · 2025Article
- Toward Antibody Production in Genome-MinimizedACS synthetic biology · 2025Article
- Establishment of the CRISPR-Cpf1 gene editing system inSynthetic and systems biotechnology · 2025Article
- Protein Engineering of Vip3A in a Selected Bacillus thuringiensis Host for Consistent High Protein Production.Current microbiology · 2024Article
- Surface-associated residues in subtilisins contribute to poly-L-lactic acid depolymerization via enzyme adsorption.Microbial biotechnology · 2024Article
- Exploring the potential of Bacillus subtilis as cell factory for food ingredients and special chemicals.Microbial cell factories · 2023Article
- Efficient production of α-monoglucosyl hesperidin by cyclodextrin glucanotransferase from Bacillus subtilis.Applied microbiology and biotechnology · 2023Article
- Linocin M18 protein from the insect pathogenic bacterium Brevibacillus laterosporus isolates.Applied microbiology and biotechnology · 2023Article
- A host-vector toolbox for improved secretory protein overproduction in Bacillus subtilis.Applied microbiology and biotechnology · 2022Article
- High antimicrobial activity of lactoferricin-expressing Bacillus subtilis strains.Microbial biotechnology · 2022Article
- UsingMarine drugs · 2021Article
- A Type I Restriction Modification System Influences Genomic Evolution Driven by Horizontal Gene Transfer inFrontiers in microbiology · 2021Article
- An efficient ABC transporter signal peptide directs heterologous protein secretion in food-grade hosts.World journal of microbiology & biotechnology · 2020Article
- Impact of spatial proximity on territoriality among human skin bacteria.NPJ biofilms and microbiomes · 2020Article
15 more citing papers are in PubMed but not listed here.
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Authors and funding
7 authors at 1 institution in 1 country.
Funding
No grant is acknowledged in the PubMed record.
Abstract
To develop an ideal blood clot imaging and targeting agent, a single-chain antibody (SCA) fragment based on a fibrin-specific monoclonal antibody, MH-1, was constructed and produced via secretion from Bacillus subtilis. Through a systematic study involving a series of B. subtilis strains, insufficient intracellular and extracytoplasmic molecular chaperones and high sensitivity to wall-bound protease (WprA) were believed to be the major factors that lead to poor production of MH-1 SCA. Intracellular and extracytoplasmic molecular chaperones apparently act in a sequential manner. The combination of enhanced coproduction of both molecular chaperones and wprA inactivation leads to the development of an engineered B. subtilis strain, WB800HM[pEPP]. This strain allows secretory production of MH-1 SCA at a level of 10 to 15 mg/liter. In contrast, with WB700N (a seven-extracellular-protease-deficient strain) as the host, no MH-1 SCA could be detected in both secreted and cellular fractions. Secreted MH-1 SCA from WB800HM[pMH1, pEPP] could be affinity purified using a protein L matrix. It retains comparable affinity and specificity as the parental MH-1 monoclonal antibody. This expression system can potentially be applied to produce other single-chain antibody fragments, especially those with folding and protease sensitivity problems.
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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.