Production of single chain Fab (scFab) fragments in Bacillus megaterium
| Autor: | Michael Hust, Stefan Dübel, Thomas Schirrmann, Eva Jordan, Laila Al-Halabi |
|---|---|
| Jazyk: | angličtina |
| Rok vydání: | 2007 |
| Předmět: |
lcsh:QR1-502
Bioengineering Single chain medicine.disease_cause Applied Microbiology and Biotechnology lcsh:Microbiology law.invention Microbiology law Fab Fragments medicine Escherichia coli Bacillus megaterium biology Chemistry Research fungi biology.organism_classification Antigen binding Biochemistry biology.protein Recombinant DNA bacteria Antibody Bacteria Biotechnology |
| Zdroj: | Microbial Cell Factories Microbial Cell Factories, Vol 6, Iss 1, p 38 (2007) |
| ISSN: | 1475-2859 |
| Popis: | BackgroundThe demand on antigen binding reagents in research, diagnostics and therapy raises questions for novel antibody formats as well as appropriate production systems. Recently, the novel single chain Fab (scFab) antibody format combining properties of single chain Fv (scFv) and Fab fragments was produced in the Gram-negative bacteriumEscherichia coli. In this study we evaluated the Gram-positive bacteriumBacillus megateriumfor the recombinant production of scFab and scFvs in comparison toE. coli.ResultsThe lysozyme specific D1.3 scFab was produced inB. megateriumandE. coli. The total yield of the scFab after purification obtained from the periplasmic fraction and culture supernatant ofE. coliwas slightly higher than that obtained from culture supernatant ofB. megaterium. However, the yield of functional scFab determined by analyzing the antigen binding activity was equally in both production systems. Furthermore, a scFv fragment with specificity for the human C reactive protein was produced inB. megaterium. The total yield of the anti-CRP scFv produced inB. megateriumwas slightly lower compared toE. coli, whereas the specific activity of the purified scFvs produced inB. megateriumwas higher compared toE. coli.ConclusionB. megateriumallows the secretory production of antibody fragments including the novel scFab antibody format. The yield and quality of functional antibody fragment is comparable to the periplasmic production inE. coli. |
| Databáze: | OpenAIRE |
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