title: Human brain pyridoxal-5'-phosphate phosphatase: production and characterization of monoclonal antibodies.
abstract: We cloned and expressed human pyridoxal-5\'-phosphate (PLP) phosphatase, the coenzymatically active form of vitamin B6, 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 B6 abnormalities.
authors.affiliation: Department of Biomedical Science and Research Institute for Bioscience and Biotechnology, Hallym University, Chunchon 200-702, Korea.
authors.forename: Dae Won
authors.initials: DW
authors.lastname: Kim
authors.forename: Won Sik
authors.initials: WS
authors.lastname: Eum
authors.forename: Hee Soon
authors.initials: HS
authors.lastname: Choi
authors.forename: So Young
authors.initials: SY
authors.lastname: Kim
authors.forename: Jae Jin
authors.initials: JJ
authors.lastname: An
authors.forename: Sun Hwa
authors.initials: SH
authors.lastname: Lee
authors.forename: Eun Joung
authors.initials: EJ
authors.lastname: Sohn
authors.forename: Seok-Il
authors.initials: SI
authors.lastname: Hwang
authors.forename: Oh-Shin
authors.initials: OS
authors.lastname: Kwon
authors.forename: Tae-Cheon
authors.initials: TC
authors.lastname: Kang
authors.forename: Moo Ho
authors.initials: MH
authors.lastname: Won
authors.forename: Sung-Woo
authors.initials: SW
authors.lastname: Cho
authors.forename: Kil Soo
authors.initials: KS
authors.lastname: Lee
authors.forename: Jinseu
authors.initials: J
authors.lastname: Park
authors.forename: Soo Young
authors.initials: SY
authors.lastname: Choi
mesh_terms: D000818:Animals; D000911:Antibodies, Monoclonal / Q000737:chemistry*; D001671:Biochemistry / Q000379:methods; D015374:Biosensing Techniques; D001921:Brain / Q000201:enzymology*; D018076:DNA, Complementary / Q000378:metabolism; D018604:Epitope Mapping; D000939:Epitopes / Q000737:chemistry; D004926:Escherichia coli / Q000378:metabolism; D015723:Gene Library; D005822:Genetic Vectors; D006801:Humans; D007150:Immunohistochemistry / Q000379:methods; D010744:Phosphoric Monoester Hydrolases / Q000737:chemistry* / Q000378:metabolism*; D051381:Rats
chemical_list: D000911:Antibodies, Monoclonal; D018076:DNA, Complementary; D000939:Epitopes; C022493:pyridoxal-5-phosphatase; D010744:Phosphoric Monoester Hydrolases