Identification by phage display of single-domain antibody fragments specific for the ODD domain in hypoxia-inducible factor 1alpha

Arjan J. Groot, Peter Verheesen, Elike J. Westerlaken, Eelke H. Gort, Petra Van Der Groep, Niels Bovenschen, Elsken Van Der Wall, Paul J. Van Diest, Avi Shvarts

Research output: Contribution to journalArticle

Abstract

Hypoxia triggers the transcription of genes responsible for cell survival via the key player transcription factor hypoxia-inducible factor 1 alpha (HIF-1α). Overexpression of this protein has been implicated in cardiovascular disorders, carcinogenesis and cancer progression. For functional and diagnostic studies on the HIF-1α protein, we have identified single-domain antibody fragments directed against this protein by using a llama-derived nonimmune phage display library. This library displays the variable domains of the heavy-chain antibody subclass, found in these animals. Phage display selection with six recombinant HIF-1α proteins yielded five different antibody fragments. By epitope-mapping, we show that all five antibody fragments bind within the functionally important oxygen-dependent degradation domain of the HIF-1α protein. Two of these antibody fragments were engineered into bivalent antibodies that were able to detect human HIF-1α by immunohistochemistry, Western blotting and immunoprecipitation, and mouse HIF-1α by immunofluorescence and immunoprecipitation. These are the first single-domain antibody fragments that may be used in exploration of HIF-1α as a possible therapeutic target through molecular applications.

Original languageEnglish (US)
Pages (from-to)345-356
Number of pages12
JournalLaboratory Investigation
Volume86
Issue number4
DOIs
Publication statusPublished - Apr 2006
Externally publishedYes

    Fingerprint

Keywords

  • Cancer
  • Hetero-bivalent VHH
  • HIF-1α
  • Hypoxia
  • Phage display
  • Single-domain antibody fragment
  • VHH

ASJC Scopus subject areas

  • Pathology and Forensic Medicine

Cite this

Groot, A. J., Verheesen, P., Westerlaken, E. J., Gort, E. H., Van Der Groep, P., Bovenschen, N., ... Shvarts, A. (2006). Identification by phage display of single-domain antibody fragments specific for the ODD domain in hypoxia-inducible factor 1alpha. Laboratory Investigation, 86(4), 345-356. https://doi.org/10.1038/labinvest.3700395