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 Citations (Scopus)

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 1 2006

Fingerprint

Hydroxymethyl and Formyl Transferases
Monoclonal Antibodies
glycinamide ribonucleotide
Enzymes
Folic Acid
Non-Small Cell Lung Carcinoma
Sodium Dodecyl Sulfate
Epitopes
Colon
Proteins
Adenocarcinoma
Immunoglobulin G
Western Blotting
Immunohistochemistry

ASJC Scopus subject areas

  • Immunology and Allergy
  • Immunology

Cite this

Dotzlaf, J., Carpenter, J., Luo, S., Roberts, E. F., Solenberg, P. J., Qian, Y. W., ... 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

Derivation and characterization of a monoclonal antibody against human glycinamide ribonucleotide formyltransferase. / Dotzlaf, Joe; Carpenter, John; Luo, Shuang; Roberts, Edda F.; Solenberg, Patricia J.; Qian, Yue Wei; Lin, Aimin; He, Xiaohua; Sandusky, George E.; McClure, Don B.; Chen, Victor J.; Zuckerman, Steven H.

In: Hybridoma, Vol. 25, No. 3, 01.06.2006, p. 139-144.

Research output: Contribution to journalArticle

Dotzlaf, J, Carpenter, J, Luo, S, Roberts, EF, Solenberg, PJ, Qian, YW, Lin, A, He, X, Sandusky, GE, McClure, DB, Chen, VJ & Zuckerman, SH 2006, 'Derivation and characterization of a monoclonal antibody against human glycinamide ribonucleotide formyltransferase', Hybridoma, vol. 25, no. 3, pp. 139-144. https://doi.org/10.1089/hyb.2006.25.139
Dotzlaf, Joe ; Carpenter, John ; Luo, Shuang ; Roberts, Edda F. ; Solenberg, Patricia J. ; Qian, Yue Wei ; Lin, Aimin ; He, Xiaohua ; Sandusky, George E. ; McClure, Don B. ; Chen, Victor J. ; Zuckerman, Steven H. / Derivation and characterization of a monoclonal antibody against human glycinamide ribonucleotide formyltransferase. In: Hybridoma. 2006 ; Vol. 25, No. 3. pp. 139-144.
@article{4a8fe3c795624f769e81d4c54d87323e,
title = "Derivation and characterization of a monoclonal antibody against human glycinamide ribonucleotide formyltransferase",
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.",
author = "Joe Dotzlaf and John Carpenter and Shuang Luo and Roberts, {Edda F.} and Solenberg, {Patricia J.} and Qian, {Yue Wei} and Aimin Lin and Xiaohua He and Sandusky, {George E.} and McClure, {Don B.} and Chen, {Victor J.} and Zuckerman, {Steven H.}",
year = "2006",
month = "6",
day = "1",
doi = "10.1089/hyb.2006.25.139",
language = "English (US)",
volume = "25",
pages = "139--144",
journal = "Monoclonal Antibodies in Immunodiagnosis and Immunotherapy",
issn = "2167-9436",
publisher = "Mary Ann Liebert Inc.",
number = "3",

}

TY - JOUR

T1 - Derivation and characterization of a monoclonal antibody against human glycinamide ribonucleotide formyltransferase

AU - Dotzlaf, Joe

AU - Carpenter, John

AU - Luo, Shuang

AU - Roberts, Edda F.

AU - Solenberg, Patricia J.

AU - Qian, Yue Wei

AU - Lin, Aimin

AU - He, Xiaohua

AU - Sandusky, George E.

AU - McClure, Don B.

AU - Chen, Victor J.

AU - Zuckerman, Steven H.

PY - 2006/6/1

Y1 - 2006/6/1

N2 - 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.

AB - 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.

UR - http://www.scopus.com/inward/record.url?scp=33745640924&partnerID=8YFLogxK

UR - http://www.scopus.com/inward/citedby.url?scp=33745640924&partnerID=8YFLogxK

U2 - 10.1089/hyb.2006.25.139

DO - 10.1089/hyb.2006.25.139

M3 - Article

C2 - 16796460

AN - SCOPUS:33745640924

VL - 25

SP - 139

EP - 144

JO - Monoclonal Antibodies in Immunodiagnosis and Immunotherapy

JF - Monoclonal Antibodies in Immunodiagnosis and Immunotherapy

SN - 2167-9436

IS - 3

ER -