Derivation and characterization of a monoclonal antibody against human glycinamide ribonucleotide formyltransferase

Joe Dotzlaf, John Carpenter, Shuang Luo, Edda F. Roberts, Patricia J. Solenberg, Yue Wei Qian, Aimin Lin, Xiaohua He, George E. Sandusky, Don B. McClure, Victor J. Chen, Steven H. Zuckerman

Research output: Contribution to journalArticle

3 Scopus citations

Abstract

Glycinamide ribonucleotide formyltransferase (GARFT) is a trifunctional enzyme involved in purine biosynthesis. Its central role in folate metabolism has made it an obvious target for the development of GARFT inhibitors, primarily for oncology. While the crystal structure, enzyme kinetics, and mechanism of action of GARFT inhibitors are reasonably well understood, GARFT regulation at the protein level remains unclear. The present study reports the development and characterization of a monoclonal antibody (MAb) specific for human GARFT. This MAb, an IgG1κ, designated PHR1, recognizes human GARFT by both Western blot and by immunohistochemistry from non-small-cell lung carcinoma and colon adenocarcinoma tissue biopsies, has a KD of 1.14 × 10-10 M, and has been epitope mapped at residues 59-78 of the GARFT functional domain. The ability of PHR1 to recognize both sodium dodecyl sulfate (SBS)-denatwred as well as native GARFT should make this MAb an important research tool in determining GARFT protein levels in both normal and neoplastic tissues.

Original languageEnglish (US)
Pages (from-to)139-144
Number of pages6
JournalHybridoma
Volume25
Issue number3
DOIs
StatePublished - Jun 2006
Externally publishedYes

ASJC Scopus subject areas

  • Immunology and Allergy
  • Immunology

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    Dotzlaf, J., Carpenter, J., Luo, S., Roberts, E. F., Solenberg, P. J., Qian, Y. W., Lin, A., He, X., Sandusky, G. E., McClure, D. B., Chen, V. J., & Zuckerman, S. H. (2006). Derivation and characterization of a monoclonal antibody against human glycinamide ribonucleotide formyltransferase. Hybridoma, 25(3), 139-144. https://doi.org/10.1089/hyb.2006.25.139