A novel nanobody against urease activity of Helicobacter pylori
| Autor: | Hamid Bakherad, Seyed Latif Mousavi Gargari, Masoumeh Rajabi Bazl, Walead Ebrahimizadeh, Iraj Rasooli, Leila Safaee Ardekani, Hamed Zare, Mohammad Taghi Mohammadi |
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| Rok vydání: | 2013 |
| Předmět: |
Microbiology (medical)
Camelid Phage display Phagemid Antibody Affinity Cross Reactions Biology Heavy chain antibody Microbiology law.invention law Animals Humans Polyacrylamide gel electrophoresis Gene Library Helicobacter pylori Heavy-chain antibody Protein Stability Temperature General Medicine Single-Domain Antibodies biology.organism_classification Antibodies Bacterial Antibodies Neutralizing Urease Molecular biology Blot Infectious Diseases Proteolysis Nanobody biology.protein Recombinant DNA Antibody Protein Binding |
| Zdroj: | International Journal of Infectious Diseases. 17:e723-e728 |
| ISSN: | 1201-9712 |
| DOI: | 10.1016/j.ijid.2013.02.015 |
| Popis: | Summary Background Helicobacter pylori infection is associated with gastritis and in some cases with gastric and duodenal ulcers, and even adenocarcinoma. Antibiotic therapy has significant limitations, such as the high cost and the emergence of antibiotic-resistant strains, generating the need for new treatments. The administration of antibody against H. pylori is a new effective therapeutic strategy. In this study, we successfully developed a single-variable domain of heavy chain antibody against recombinant UreC. Methods A VHH phagemid library was constructed from immune camel heavy chain antibodies. The nanobodies were displayed on M13 phage. Library selection was performed against UreC recombinant protein. A specific single-variable domain of heavy chain antibody against UreC was screened in five rounds of panning. The nanobody with the highest score in the phage ELISA was selected for soluble expression. The nanobody was purified with a nickel–nitrilotriacetic acid (Ni–NTA) column and confirmed with sodium dodecyl sulfate polyacrylamide gel electrophoresis (SDS-PAGE) and Western blotting. Affinity, specificity, and urease inhibitory properties of the nanobody were assayed. Results Here we showed the isolation and purification of a specific nanobody with high affinity against UreC recombinant protein that can inhibit urease activity. Conclusions The isolated UreC nanobody can specifically detect and bind to UreC and inhibit urease activity. This nanobody could be a novel class of treatment measure against H. pylori infection. |
| Databáze: | OpenAIRE |
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