Preparation of anti-orotate phosphoribosyltransferase antibody and its application to immunochemical detection in human tumor cells.
Orotate phosphoribosyltransferase (OPRT) is a key enzyme in the anabolism of 5-fluorouracil (5-FU), and its expression in tumors is thought to increase the efficacy of 5-FU against the tumor. To detect the OPRT protein by immunoblot and/or immunohistochemical methods, we prepared highly specific antibody to the peptides contained in the human OPRT amino acid sequence. The anti-OPRT polyclonal antibodies, obtained by immunizing rabbits with the OPRT peptides, had a high specificity for the OPRT protein in human tumor xenografts when it was analyzed by immunoblotting, and furthermore, there was a positive correlation between OPRT activity and protein content in 12 human tumors (R2 = 0.632). These results suggest that immunohistochemical detection of tumoral OPRT protein expression with our anti-OPRT antibodies may provide a useful method for predicting the clinical response to 5-FU-based chemotherapy.
Sakamoto
Kazuki
K
Optimal Medication Research Laboratory, Taiho Pharmaceutical Co., Ltd., 224-2 Hiraishi-Ebisuno, Tokushima 771-0194, Japan. ka-sakamoto@taiho.co.jp
Sugimoto
Yoshikazu
Y
Miyadera
Kazutaka
K
Oka
Toshinori
T
Fukushima
Masakazu
M
D000595:Amino Acid Sequence; D000818:Animals; D000911:Antibodies, Monoclonal / Q000276:immunology* / Q000302:isolation & purification; D000915:Antibody Affinity / Q000276:immunology; D000918:Antibody Specificity / Q000276:immunology; D045744:Cell Line, Tumor; D004797:Enzyme-Linked Immunosorbent Assay; D005354:Fibrosarcoma / Q000276:immunology* / Q000473:pathology; D006801:Humans; D015151:Immunoblotting; D007150:Immunohistochemistry; D051379:Mice; D008807:Mice, Inbred BALB C; D008819:Mice, Nude; D008856:Microscopy, Fluorescence; D008969:Molecular Sequence Data; D009374:Neoplasms, Experimental / Q000276:immunology / Q000378:metabolism / Q000473:pathology; D009962:Orotate Phosphoribosyltransferase / Q000235:genetics / Q000276:immunology* / Q000378:metabolism; D011817:Rabbits; D014183:Transplantation, Heterologous
D000911:Antibodies, Monoclonal; D009962:Orotate Phosphoribosyltransferase