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Monoclonal Antibody-Based Screening Assay for Factor-Inhibiting Hypoxia-Inducible Factor Inhibitors
Korea Research Institute of Bioscience and Biotechnology
* To whom correspondence should be addressed. E-mail: mklee{at}kribb.re.kr.
 (HIF-1), and the modification abrogates the transcriptional activity of HIF-1. Because FIH is more active on HIF1 than prolyl hydroxylase domain proteins under hypoxic conditions, its inhibitors have potential to be developed as anti-ischemic drugs targeting normal cells stressed by hypoxia. In this study, the authors developed the first monoclonal antibody, SHN-HIF1, specifically to Asn803 hydroxylated HIF-1 and a sensitive assay system for FIH inhibitors using the monoclonal antibody (Mab). SHN-HIF1 showed 740 times higher affinity to the Asn803 hydroxylated HIF-1 peptide than the unmodified one. An enzyme-linked immunosorbent assay–based system using SHN-HIF1 displayed at least 30 times more sensitivity than previous methods for screening FIH inhibitors and was easily applicable to develop a high-throughput screening system. SHNHIF1 also showed an Asn803 hydroxylation-dependent specificity to HIF-1 in cells. Taken together, the results suggest that it may be used to analyze the in vivo and in vitro activities of FIH inhibitors. (Journal of Biomolecular Screening XXXX:xx-xx)
First published on June 19, 2008, doi:10.1177/1087057108318800 This article has been cited by other articles:
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