- 作者：Leila Safaee Ardekani ; Seyed Latif Mousavi Gargari ; Iraj Rasooli ; Masoumeh Rajabi Bazl ; Mohammad Mohammadi ; Walead Ebrahimizadeh ; Hamid Bakherad ; Hamed Zare
- 关键词：Helicobacter pylori ; Phage display ; Camelid ; Heavy chain antibody ; Nanobody
- 刊名：International Journal of Infectious Diseases
- 出版年：2013
- 出版时间：September, 2013
- 年：2013
- 卷：17
- 期：9
- 页码：e723-e728
- 全文大小：853 K
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.