Preparation of anti-orotate phosphoribosyltransferase antibody and its application to immunochemical detection in human tumor cells.

Abstract

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.

• Authors: Kazuki Sakamoto, Yoshikazu Sugimoto, Kazutaka Miyadera, Toshinori Oka, Masakazu Fukushima
• Journal: International journal of molecular medicine (Int J Mol Med) Vol. 16 Issue 2 Pg. 245-9 (Aug 2005) ISSN: 1107-3756 [Print] Greece
• PMID: 16012756 (Publication Type: Journal Article)

Chemical References

• Antibodies, Monoclonal
• Orotate Phosphoribosyltransferase

Topics

• Amino Acid Sequence
• Animals
• Antibodies, Monoclonal (immunology, isolation & purification)
• Antibody Affinity (immunology)
• Antibody Specificity (immunology)
• Cell Line, Tumor
• Enzyme-Linked Immunosorbent Assay
• Fibrosarcoma (immunology, pathology)
• Humans
• Immunoblotting
• Immunohistochemistry
• Mice
• Mice, Inbred BALB C
• Mice, Nude
• Microscopy, Fluorescence
• Molecular Sequence Data
• Neoplasms, Experimental (immunology, metabolism, pathology)
• Orotate Phosphoribosyltransferase (genetics, immunology, metabolism)
• Rabbits
• Transplantation, Heterologous