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cDNA‐directed expression of human thyroid peroxidase
Author(s) -
Kimura Shioko,
Kotani Tomio,
Ohtaki Sachiya,
Aoyama Toshifumi
Publication year - 1989
Publication title -
febs letters
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.593
H-Index - 257
eISSN - 1873-3468
pISSN - 0014-5793
DOI - 10.1016/0014-5793(89)80759-8
Subject(s) - complementary dna , peroxidase , microbiology and biotechnology , thyroid peroxidase , monoclonal antibody , affinity chromatography , guaiacol , biology , chemistry , antibody , biochemistry , enzyme , gene , immunology
A human thyroid peroxidase cDNA, hTPO‐1 [(1987) Proc. Natl. Acad. Sci. USA 84, 5555–5559], was expressed in human Hep G2 cells using a vaccinia virus cDNA‐expression system. When examined by immunoblot analysis, the level of hTPO‐1 protein expression reached a maximum approx. 24 h after infection and remained at a similar level up to 72 h post‐infection. The expressed protein was enzymatically active as measured by guaiacol oxidation. Monoclonal antibody‐assisted immunoaffinity column chromatography was used for partial purification of vaccinia‐expressed hTPO‐1, resulting in more than 300‐fold higher specific activity and a measurable difference spectrum of the hTPO‐1 (Fe 3+ )‐CN complex.