Derivation and characterization of a monoclonal antibody against human glycinamide ribonucleotide formyltransferase.
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 IgG1kappa, 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 x 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 (SDS)-denatured as well as native GARFT should make this MAb an important research tool in determining GARFT protein levels in both normal and neoplastic tissues.
Dotzlaf
Joe
J
Lilly Research Laboratories, Eli Lilly & Company, Indianapolis, Indiana 46285, USA.
Carpenter
John
J
Luo
Shuang
S
Roberts
Edda F
EF
Solenberg
Patricia J
PJ
Qian
Yue-Wei
YW
Lin
Aimin
A
He
Xiaohua
X
Sandusky
George E
GE
McClure
Don B
DB
Chen
Victor J
VJ
Zuckerman
Steven H
SH
D000595:Amino Acid Sequence; D000818:Animals; D000911:Antibodies, Monoclonal / Q000737:chemistry; D000939:Epitopes / Q000235:genetics / Q000276:immunology; D006367:HeLa Cells; D006801:Humans; D006825:Hybridomas; D051379:Mice; D008807:Mice, Inbred BALB C; D008969:Molecular Sequence Data; D051026:Phosphoribosylglycinamide Formyltransferase / Q000235:genetics / Q000276:immunology*
D000911:Antibodies, Monoclonal; D000939:Epitopes; D051026:Phosphoribosylglycinamide Formyltransferase