RNAi suppression of β-N-acetylglucosaminidase (BmFDL) for complex-type N-linked glycan synthesis in cultured silkworm cells

Yudai Nagata, Man Lee, Hiroaki Mon, Shigeo Imanishi, Sun Mee Hong, Shoji Komatsu, Yuji Oshima, Takahiro Kusakabe

Research output: Contribution to journalArticle

14 Citations (Scopus)

Abstract

Glycoproteins have various biological functions including enzymatic activity, protein stability and others. Due to the presence of paucimannosidic N-linked glycans, recombinant proteins from an insect cell expression system may not be suitable for therapeutic use. Because baculovirus expression systems (BESs) are used to produce recombinant proteins, it is of interest to modify the endogenous N-glycosylation pathway in insects to mimic that of mammals. Using a soaking RNAi sensitive cell line, BmN4-SID1, has enabled us to suppress Bombyx mori FDL (BmFDL), an N-linked glycan-specific β-N-acetylglucosaminidase. Western blotting and MALDI-TOF MS demonstrated that the BmFDL depletion almost completely converted the paucimannosidic structures of the recombinant proteins produced by BES into a complex-type structure. This highly efficient, simple and low-cost method can be used for mass production of secretion proteins with complex-type N-linked glycans.

Original languageEnglish
Pages (from-to)1009-1016
Number of pages8
JournalBiotechnology letters
Volume35
Issue number7
DOIs
Publication statusPublished - Jul 1 2013

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Recombinant proteins
Acetylglucosaminidase
Bombyx
RNA Interference
Recombinant Proteins
Polysaccharides
Cultured Cells
Baculoviridae
Insects
Glycosylation
Proteins
Glycoproteins
Mammals
Protein Stability
Matrix-Assisted Laser Desorption-Ionization Mass Spectrometry
Therapeutic Uses
Western Blotting
Cells
Costs and Cost Analysis
Cell Line

All Science Journal Classification (ASJC) codes

  • Biotechnology
  • Bioengineering
  • Applied Microbiology and Biotechnology

Cite this

RNAi suppression of β-N-acetylglucosaminidase (BmFDL) for complex-type N-linked glycan synthesis in cultured silkworm cells. / Nagata, Yudai; Lee, Man; Mon, Hiroaki; Imanishi, Shigeo; Hong, Sun Mee; Komatsu, Shoji; Oshima, Yuji; Kusakabe, Takahiro.

In: Biotechnology letters, Vol. 35, No. 7, 01.07.2013, p. 1009-1016.

Research output: Contribution to journalArticle

Nagata, Yudai ; Lee, Man ; Mon, Hiroaki ; Imanishi, Shigeo ; Hong, Sun Mee ; Komatsu, Shoji ; Oshima, Yuji ; Kusakabe, Takahiro. / RNAi suppression of β-N-acetylglucosaminidase (BmFDL) for complex-type N-linked glycan synthesis in cultured silkworm cells. In: Biotechnology letters. 2013 ; Vol. 35, No. 7. pp. 1009-1016.
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AU - Hong, Sun Mee

AU - Komatsu, Shoji

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