[Preparation of anti-OPRT antibody for immunochemical detection].
Orotate phosphoribosyl transferase (OPRT, EC 2.4.2.10) is a key enzyme in the anabolism of 5-fluorouracil (5 FU), and its expression in tumor is thought to increase the efficacy of 5-FU against the tumor. To detect the OPRT protein by immunoblotting and/or immunohistochemical methods, we tried to prepare highly specific antibody against the peptide including human OPRT amino acid sequence. The anti-OPRT polyclonal antibody obtained by immunization of OPRT peptides to rabbits had high specificity for the OPRT protein in human tumor xenografts in immunoblot and immunohistochemical analysis. Furthermore, there was a positive correlation between the activity and proteins of OPRT (R2=0.632) in 12 human tumors. These results suggest that immunohistochemical detection of tumoral OPRT protein expression using our anti-OPRT antibody may provide useful methods to predict the clinical response to 5-FU-based chemotherapy.
Sakamoto
Kazuki
K
The Optimal Medication Research Laboratory, Taiho Pharmaceutical Co., Ltd.
Sugimoto
Yoshikazu
Y
Miyadera
Kazutaka
K
Oka
Toshinori
T
Fukushima
Masakazu
M
D000595:Amino Acid Sequence; D000818:Animals; D000911:Antibodies, Monoclonal / Q000096:biosynthesis*; D005354:Fibrosarcoma / Q000201:enzymology*; D005472:Fluorouracil / Q000378:metabolism; D006801:Humans; D015151:Immunoblotting; D007150:Immunohistochemistry; D051379:Mice; D008819:Mice, Nude; D009368:Neoplasm Transplantation; D009962:Orotate Phosphoribosyltransferase / Q000737:chemistry / Q000276:immunology* / Q000378:metabolism
D000911:Antibodies, Monoclonal; D009962:Orotate Phosphoribosyltransferase; D005472:Fluorouracil