Production of biologically active scFv and VHH antibody fragments in Bifidobacterium longum

Bifidobacteria constitute a significant part of healthy intestinal microbiota in adults and infants and present a promising platform for construction of genetically modified probiotic agents for treatment of gastrointestinal disorders. In this study, three strains of Bifidobacterium longum were cons...

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Bibliographic Details
Published in:FEMS microbiology letters 2015-06, Vol.362 (12), p.fnv083-fnv083
Main Authors: Shkoporov, A. N., Khokhlova, E. V., Savochkin, K. A., Kafarskaia, L. I., Efimov, B. A.
Format: Article
Language:English
Subjects:
Adalimumab - biosynthesis
Animals
Antibodies
Antibodies, Bacterial - biosynthesis
Bifidobacterium - genetics
Bifidobacterium - metabolism
Bifidobacterium longum
Biological activity
Cell culture
Cells, Cultured
Cytoplasm
Endotoxins - metabolism
Exotoxin A
Gastrointestinal diseases
Gene Expression
Genetic modification
Humans
Infants
Inflammatory bowel diseases
Intestinal microflora
Intestine
Male
Mice, Inbred C57BL
Mice, Inbred DBA
Microbiology
Microbiota
Organisms, Genetically Modified
Probiotics
Tumor necrosis factor-α
Yogurt
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Summary:Bifidobacteria constitute a significant part of healthy intestinal microbiota in adults and infants and present a promising platform for construction of genetically modified probiotic agents for treatment of gastrointestinal disorders. In this study, three strains of Bifidobacterium longum were constructed that express and secrete biologically active single-chain antibodies against human TNF-α and Clostridium difficile exotoxin A. Anti-TNF-α scFv antibody D2E7 was produced at the level of 25 μg L−1 in broth culture and was mostly retained in the cytoplasm, while VHH-type antibodies A20.1 and A26.8 against C. difficile exotoxin A were produced at the levels of 0.3–1 mg L−1 and secreted very efficiently. The biological activity of both antibody types was demonstrated in the mammalian cell-based assays. Expression of A20.1 and A26.8 was also observed in vivo after intragastric administration of transformed B. longum strains to (C57/BL6 × DBA/2)F1 mice. The obtained B. longum strains may serve as prototypes for construction of novel probiotic medications against inflammatory bowel disease and C. difficile-associated disease. Three strains of bifidobacteria were constructed that secrete antibodies against human TNF-alpha and Clostridium difficile exotoxin TcdA.
ISSN:1574-6968
0378-1097
1574-6968
DOI:10.1093/femsle/fnv083