Selection of a fully human single domain antibody specific to Helicobacter pylori urease

Applied Microbiology and Biotechnology
Mehdi FouladiYadollah Omidi

Abstract

Helicobacter pylori bacteria are involved in gastroduodenal disorders, including gastric adenocarcinoma. Since the current therapies encounter with some significant shortcomings, much attention has been paid to the development of new alternative diagnostic and treatment modalities such as immunomedicines to target H. pylori. Having used phage display technology, we isolated fully humane small antibody (Ab) fragment (VL) against the Flap region of urease enzyme of H. pylori to suppress its enzymatic activity. Solution biopanning (SPB) and screening process against a customized biotinylated peptide corresponding to the enzyme Flap region resulted in the selection of VL single domain Abs confirmed by the enzyme-linked immunosorbent assay (ELISA), sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE), and Western blotting. The selected Ab fragments showed a high affinity with a KD value of 97.8 × 10-9 and specificity to the enzyme with high inhibitory impact. For the first time, a VL single domain Ab was isolated by SPB process against a critical segment of H. pylori urease using a diverse semi-synthetic library. Based on our findings, the selected VL Ab fragments can be used for the diagnosis, imaging, targeting, an...Continue Reading

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Citations

Sep 10, 2020·International Journal of Peptide Research and Therapeutics·Chunyan LiHong Yang
Oct 8, 2020·Journal of Translational Medicine·Tao ChenHenry Wei
Dec 22, 2019·Biotechnology Advances·Jaber DehghaniYadollah Omidi
Jul 27, 2021·Frontiers in Cellular and Infection Microbiology·Kristian Daniel Ralph RothMichael Hust

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Datasets Mentioned

BETA
LC375193

Methods Mentioned

BETA
enzyme-linked
enzyme-linked immunosorbent assay
ELISA
electrophoresis
X-ray
surface plasmon resonance
biopsy
PCR
Surface
phage display

Software Mentioned

Frustratometer
ProsA
LOMETS
VBASE2
SPICKER
PSSpred
TIGRESS
Verify
verify3D
YASARA

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