Selection by phage display of nanobodies directed against hypoxia inducible factor-1α (HIF-1α).

Abstract

Hypoxia, which promotes tumor invasion and metastasis, is a common phenomenon in solid tumors. Hypoxia generally leads to a higher expression level of hypoxia inducible factor-1 (HIF-1) in tumors (cells) relative to normal tissues (cells). Given the unique expression of HIF-1α in human cancers and its vital importance in mediating hypoxic adaptation, we have identified 20 different HIF-1α-specific nanobodies by using a llama-derived nonimmune phage display library. PAS-B domain of HIF-1α (HIF-1α-PAS-B) has been used as an antigen. Nanobody (VHH16) was selected from these 20 nanobodies by phage enzyme-linked immunosorbent assay. The preliminary analysis of biological activity demonstrates that VHH16 can specifically bind to HIF-1α with high affinity. VHH16 is the first nanobody that specifically binds to HIF-1α-PAS-B as well. We suggest here that VHH16 is useful in disease diagnosis and also has potential in medical applications.

• Authors: Min Li, Xiaodan Fan, Jing Liu, Yaozhong Hu, He Huang
• Journal: Biotechnology and applied biochemistry (Biotechnol Appl Biochem) 2015 Nov-Dec Vol. 62 Issue 6 Pg. 738-45 ISSN: 1470-8744 [Electronic] United States
• PMID: 25556956 (Publication Type: Journal Article, Research Support, Non-U.S. Gov't)
• Copyright: © 2014 International Union of Biochemistry and Molecular Biology, Inc.

Chemical References

• Hypoxia-Inducible Factor 1, alpha Subunit
• Peptide Library
• Single-Domain Antibodies

Topics

• Amino Acid Sequence
• Animals
• Antibody Specificity
• Camelids, New World
• Hypoxia-Inducible Factor 1, alpha Subunit (chemistry, immunology)
• Molecular Sequence Data
• Peptide Library
• Protein Engineering (methods)
• Protein Structure, Tertiary
• Single-Domain Antibodies (immunology)