Chapter title |
Engineering N-Glycosylation Pathway in Insect Cells: Suppression of β-N-Acetylglucosaminidase and Expression of β-1,4-Galactosyltransferase.
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Chapter number | 13 |
Book title |
Glyco-Engineering
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Published in |
Methods in molecular biology, January 2015
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DOI | 10.1007/978-1-4939-2760-9_13 |
Pubmed ID | |
Book ISBNs |
978-1-4939-2759-3, 978-1-4939-2760-9
|
Authors |
Kim, Yeon Kyu, Cha, Hyung Joon, Yeon Kyu Kim, Hyung Joon Cha |
Abstract |
Most insect cells have a simple N-glycosylation process and consequently paucimannosidic or simple core glycans predominate. It has been proposed that β-N-acetylglucosaminidase (GlcNAcase), a hexosaminidase in the Golgi membrane which removes a terminal N-acetylglucosamine (GlcNAc), might contribute to simple N-glycosylation profile in several insect cells including Drosophila S2. Here, we describe GlcNAcase suppression strategy using RNA interference (RNAi) to avoid the formation of paucimannosidic glycans in insect S2 cells. In addition, we describe coexpression of β(1,4)-galactosyltransferase (GalT) as a strategy to improve N-glycosylation pattern and enable recombinant therapeutic proteins to be produced in S2 cells with more complex N-glycans. |
Mendeley readers
Geographical breakdown
Country | Count | As % |
---|---|---|
Unknown | 9 | 100% |
Demographic breakdown
Readers by professional status | Count | As % |
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Researcher | 3 | 33% |
Student > Ph. D. Student | 2 | 22% |
Other | 1 | 11% |
Student > Master | 1 | 11% |
Unknown | 2 | 22% |
Readers by discipline | Count | As % |
---|---|---|
Agricultural and Biological Sciences | 5 | 56% |
Medicine and Dentistry | 1 | 11% |
Engineering | 1 | 11% |
Unknown | 2 | 22% |