Back to Search
Start Over
Generation and characterization of a diabody targeting the αvβ6 integrin.
- Source :
-
PloS one [PLoS One] 2013 Sep 04; Vol. 8 (9), pp. e73260. Date of Electronic Publication: 2013 Sep 04 (Print Publication: 2013). - Publication Year :
- 2013
-
Abstract
- The αvβ6 integrin is up-regulated in cancer and wound healing but it is not generally expressed in healthy adult tissue. There is increasing evidence that it has a role in cancer progression and will be a useful target for antibody-directed cancer therapies. We report a novel recombinant diabody antibody fragment that targets specifically αvβ6 and blocks its function. The diabody was engineered with a C-terminal hexahistidine tag (His tag), expressed in Pichia pastoris and purified by IMAC. Surface plasmon resonance (SPR) analysis of the purified diabody showed affinity in the nanomolar range. Pre-treatment of αvβ6-expressing cells with the diabody resulted in a reduction of cell migration and adhesion to LAP, demonstrating biological function-blocking activity. After radio-labeling, using the His-tag for site-specific attachment of (99m)Tc, the diabody retained affinity and targeted specifically to αvβ6-expressing tumors in mice bearing isogenic αvβ6 +/- xenografts. Furthermore, the diabody was specifically internalized into αvβ6-expressing cells, indicating warhead targeting potential. Our results indicate that the new αvβ6 diabody has a range of potential applications in imaging, function blocking or targeted delivery/internalization of therapeutic agents.
- Subjects :
- Animals
Antibody Specificity
Antigens, Neoplasm metabolism
Cell Line
Female
Gene Expression Regulation, Neoplastic
Humans
Immunoglobulin Fragments chemistry
Integrins metabolism
Isotope Labeling
Mice
Pichia genetics
Recombinant Proteins chemistry
Skin Neoplasms metabolism
Technetium chemistry
Antigens, Neoplasm immunology
Immunoglobulin Fragments genetics
Immunoglobulin Fragments immunology
Integrins immunology
Protein Engineering
Recombinant Proteins genetics
Recombinant Proteins immunology
Subjects
Details
- Language :
- English
- ISSN :
- 1932-6203
- Volume :
- 8
- Issue :
- 9
- Database :
- MEDLINE
- Journal :
- PloS one
- Publication Type :
- Academic Journal
- Accession number :
- 24023846
- Full Text :
- https://doi.org/10.1371/journal.pone.0073260