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Human Brain Pyridoxal-5'-phosphate Phosphatase: Production and Characterization of Monoclonal Antibodies

  • Kim, Dae-Won (Department of Anatomy, College of Medicine, Hallym University) ;
  • Eum, Won-Sik (Department of Anatomy, College of Medicine, Hallym University) ;
  • Choi, Hee-Soon (Department of Biomedical Science and Research Institute for Bioscience and Biotechnology, Hallym University) ;
  • Kim, So-Young (Department of Biomedical Science and Research Institute for Bioscience and Biotechnology, Hallym University) ;
  • An, Jae-Jin (Department of Biomedical Science and Research Institute for Bioscience and Biotechnology, Hallym University) ;
  • Lee, Sun-Hwa (Department of Biomedical Science and Research Institute for Bioscience and Biotechnology, Hallym University) ;
  • Sohn, Eun-Joung (Department of Biomedical Science and Research Institute for Bioscience and Biotechnology, Hallym University) ;
  • Hwang, Seok-Il (Department of Biomedical Science and Research Institute for Bioscience and Biotechnology, Hallym University) ;
  • Kwon, Oh-Shin (Department of Biochemistry, Kyungpook National University) ;
  • Kang, Tae-Cheon (Department of Anatomy, College of Medicine, Hallym University) ;
  • Won, Moo-Ho (Department of Anatomy, College of Medicine, Hallym University) ;
  • Cho, Sung-Woo (Department of Biochemistry and Molecular Biology, University of Ulsan College of Medicine) ;
  • Lee, Kil-Soo (Department of Biomedical Science and Research Institute for Bioscience and Biotechnology, Hallym University) ;
  • Park, Jin-Seu (Department of Biomedical Science and Research Institute for Bioscience and Biotechnology, Hallym University) ;
  • Choi, Soo-Young (Department of Biomedical Science and Research Institute for Bioscience and Biotechnology, Hallym University)
  • 발행 : 2005.11.30

초록

We cloned and expressed human pyridoxal-5'-phosphate (PLP) phosphatase, the coenzymatically active form of vitamin $B_6$, in Escherichia coli using pET15b vector. Monoclonal antibodies (mAb) were generated against purified human brain PLP phosphatase in mice, and four antibodies recognizing different epitopes were obtained, one of which inhibited PLP phosphatase. The binding affinities of these four mAbs to PLP phosphatase, as determined using biosensor technology, showed that they had similar binding affinities. Using the anti-PLP phosphatase antibodies as probes, we investigated their cross-reactivities in various mammalian and human tissues and cell lines. The immunoreactive bands obtained on Western blots had molecular masses of ca. 33 kDa. Similarly fractionated extracts of several mammalian cell lines all produced a single band of molecular mass 33 kDa. We believe that these PLP phosphatase mAbs could be used as valuable immunodiagnostic reagents for the detection, identification, and characterization of various neurological diseases related to vitamin $B_6$ abnormalities.

키워드

참고문헌

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