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Murine monoclonal antibody recognizing human α(1,3/1,4)fucosyltransferase

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Abstract

We prepared a mouse monoclonal antibody, FTA1-16, that specifically recognizes human α(1,3/1,4)fucosyltransferase without crossreactivity to any other members of the α(1,3)fucosyltransferase family. The specificity was confirmed by both immunofluorescense staining of native antigens in the Golgi apparatus and Western blotting analysis, using stable transformant cells transfected with each gene of the α(1,3)fucosyltransferase family. Western blotting analysis on a series of human tumour cell lines from various tissues revealed that some epithelial cancer cell lines from digestive organs expressed an amount of α(1,3/1,4)fucosyltransferase in good correlation with expression of sialyl Lewis a antigen. Immunohistochemical staining by FTA1-16 on colon cancer tissues revealed enhanced expression of the enzyme in cancer cells in comparison to normal cells. Finally, the antigenic epitope recognized by FTA1-16 was determined using truncated recombinant peptides which were expressed inE. coli. A minimal length determined was a fragment, amino acid positions 132–153, of the α(1,3/1,4)fucosyltransferase.

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Abbreviations

Fuc-T:

fucosyltransferase

β1,4GalT:

β1,4galactosyltransferase

mAb:

monoclonal antibody

RT-PCR:

reverse transcriptase-polymerase chain reaction

ORF:

open reading frame

PVDF:

polyvinylidene difluoride

ELISA:

enzyme linked immunosolvent assay

SDS:

sodium dodecylsulfate

PAGE:

polyacrylamide gel electrophoresis

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Authors and Affiliations

  1. Institute of Life Science, Soka University, 1-236 Tangi-cho, 192, Hachioji, Tokyo, Japan

    Hirohisa Kimura, Takashi Kudo, Shoko Nishihara, Hiroko Iwasaki, Naoko Shinya, Rihito Watanabe & Hisashi Narimatsu

  2. Department of Microbiology, Keio University School of Medicine, 35 Shinanomachi, Shinjuku-ku, 160, Tokyo, Japan

    Takashi Kudo

  3. Diagnostics Research Laboratories, Fujirebio Inc., 51 Komiya-cho, 192, Hachioji, Tokyo, Japan

    Hideo Honda & Fuminori Takemura

Authors
  1. Hirohisa Kimura
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  2. Takashi Kudo
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  3. Shoko Nishihara
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  4. Hiroko Iwasaki
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  5. Naoko Shinya
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  6. Rihito Watanabe
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  7. Hideo Honda
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  8. Fuminori Takemura
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  9. Hisashi Narimatsu
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Kimura, H., Kudo, T., Nishihara, S. et al. Murine monoclonal antibody recognizing human α(1,3/1,4)fucosyltransferase. Glycoconjugate J 12, 802–812 (1995). https://doi.org/10.1007/BF00731242

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  • DOI: https://doi.org/10.1007/BF00731242

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