Genetic engineering approach to develop next-generation reagents for endotoxin quantification

Hikaru Mizumura, Norihiko Ogura, Jun Aketagawa, Maki Aizawa, Yuki Kobayashi, Shun Ichiro Kawabata, Toshio Oda

研究成果: ジャーナルへの寄稿記事

2 引用 (Scopus)

抄録

The bacterial endotoxin test, which uses amebocyte lysate reagents of horseshoe crab origin, is a sensitive, reproducible and simple assay to measure endotoxin concentration. To develop sustainable raw materials for lysate reagents that do not require horseshoe crabs, three recombinant protease zymogens (factor C, derived from mammalian cells; factor B; and the proclotting enzyme derived from insect cells) were prepared using a genetic engineering technique. Recombinant cascade reagents (RCRs) were then prepared to reconstruct the reaction cascade in the amebocyte lysate reagent. The protease activity of the RCR containing recombinant factor C was much greater than that of recombinant factor C alone, indicating the efficiency of signal amplification in the cascade. Compared with the RCR containing the insect cell-derived factor C, those containing mammalian cell-derived factor C, which features different glycosylation patterns, were less susceptible to interference by the injectable drug components. The standard curve of the RCR containing mammalian cell-derived recombinant factor C had a steeper slope than the curves for those containing natural lysate reagents, suggesting a greater sensitivity to endotoxin. The present study supports the future production of recombinant reagents that do not require the use of natural resources.

元の言語英語
ページ(範囲)136-146
ページ数11
ジャーナルInnate Immunity
23
発行部数2
DOI
出版物ステータス出版済み - 1 1 2017

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Genetic Engineering
Endotoxins
Horseshoe Crabs
Insects
Peptide Hydrolases
Genetic Techniques
Enzyme Precursors
Complement Factor B
Glycosylation
Injections

All Science Journal Classification (ASJC) codes

  • Microbiology
  • Immunology
  • Molecular Biology
  • Cell Biology
  • Infectious Diseases

これを引用

Genetic engineering approach to develop next-generation reagents for endotoxin quantification. / Mizumura, Hikaru; Ogura, Norihiko; Aketagawa, Jun; Aizawa, Maki; Kobayashi, Yuki; Kawabata, Shun Ichiro; Oda, Toshio.

:: Innate Immunity, 巻 23, 番号 2, 01.01.2017, p. 136-146.

研究成果: ジャーナルへの寄稿記事

Mizumura, H, Ogura, N, Aketagawa, J, Aizawa, M, Kobayashi, Y, Kawabata, SI & Oda, T 2017, 'Genetic engineering approach to develop next-generation reagents for endotoxin quantification', Innate Immunity, 巻. 23, 番号 2, pp. 136-146. https://doi.org/10.1177/1753425916681074
Mizumura, Hikaru ; Ogura, Norihiko ; Aketagawa, Jun ; Aizawa, Maki ; Kobayashi, Yuki ; Kawabata, Shun Ichiro ; Oda, Toshio. / Genetic engineering approach to develop next-generation reagents for endotoxin quantification. :: Innate Immunity. 2017 ; 巻 23, 番号 2. pp. 136-146.
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