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

Kazuki Sakamoto, Yoshikazu Sugimoto, Kazutaka Miyadera, Toshinori Oka, Masakazu Fukushima

Research output: Contribution to journalArticle

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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.

Original languageEnglish
Pages (from-to)245-249
Number of pages5
JournalInternational journal of molecular medicine.
Volume16
Issue number2
Publication statusPublished - 2005 Aug
Externally publishedYes

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Orotate Phosphoribosyltransferase
Antibodies
Neoplasms
Fluorouracil
Proteins
Peptides
Immunoblotting
Heterografts
Amino Acid Sequence

ASJC Scopus subject areas

  • Genetics

Cite this

Preparation of anti-orotate phosphoribosyltransferase antibody and its application to immunochemical detection in human tumor cells. / Sakamoto, Kazuki; Sugimoto, Yoshikazu; Miyadera, Kazutaka; Oka, Toshinori; Fukushima, Masakazu.

In: International journal of molecular medicine., Vol. 16, No. 2, 08.2005, p. 245-249.

Research output: Contribution to journalArticle

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