| Item type |
文献 / Documents(1) |
| 公開日 |
2021-06-11 |
| アクセス権 |
|
|
アクセス権 |
open access |
| 資源タイプ |
|
|
資源タイプ識別子 |
http://purl.org/coar/resource_type/c_6501 |
|
資源タイプ |
journal article |
| 出版社版DOI |
|
|
|
識別子タイプ |
DOI |
|
|
関連識別子 |
https://doi.org/10.3390/biom10111482 |
|
|
言語 |
ja |
|
|
関連名称 |
10.3390/biom10111482 |
| 出版タイプ |
|
|
出版タイプ |
VoR |
|
出版タイプResource |
http://purl.org/coar/version/c_970fb48d4fbd8a85 |
| タイトル |
|
|
タイトル |
Silkworm Pupae Function as Efficient Producers of Recombinant Glycoproteins with Stable-Isotope Labeling |
|
言語 |
en |
| 著者 |
ヤギ, ヒロカズ
ヤナカ, サエコ
ヨゴ, リナ
イケダ, アカリ
鬼塚, 正義
ヤマザキ, トシオ
カトウ, タツヤ
Park, Enoch Y.
ヨコヤマ, ジュン
カトウ, コウイチ
|
| 抄録 |
|
|
内容記述タイプ |
Abstract |
|
内容記述 |
Baculovirus-infected silkworms are promising bioreactors for producing recombinant glycoproteins, including antibodies. Previously, we developed a method for isotope labeling of glycoproteins for nuclear magnetic resonance (NMR) studies using silkworm larvae reared on an artificial diet containing 15N-labeled yeast crude protein extract. Here, we further develop this method by introducing a technique for the expression of isotope-labeled glycoproteins by silkworm pupae, which has several potential advantages relative to larvae-based techniques in terms of production yield, ease of handling, and storage. Here, we fed fifth instar larvae an artificial diet with an optimized composition containing [methyl-13C]methionine, leading to pupation. Nine-day-old pupae were then injected with recombinant Bombyx mori nucleopolyhedrovirus (BmNPV) bacmid for expression of recombinant human immunoglobulin G (IgG). From the whole-body homogenates of pupae, 0.35 mg/pupa of IgG was harvested, which is a yield that is five times higher than can be obtained from larvae. Recombinant IgG, thus prepared, exhibited mainly three kinds of pauci-mannose-type oligosaccharides and had a 13C-enrichment ratio of approximately 80%. This enabled selective observation of NMR signals originating from the methionyl methyl group of IgG, confirming its conformational integrity. These data demonstrate the utility of silkworm pupae as factories for producing recombinant glycoproteins with amino-acid-selective isotope labeling. |
|
言語 |
en |
| キーワード |
|
|
言語 |
en |
|
主題Scheme |
Other |
|
主題 |
silkworm pupa |
| キーワード |
|
|
言語 |
en |
|
主題Scheme |
Other |
|
主題 |
isotope labeling |
| キーワード |
|
|
言語 |
en |
|
主題Scheme |
Other |
|
主題 |
recombinant glycoprotein |
| キーワード |
|
|
言語 |
en |
|
主題Scheme |
Other |
|
主題 |
artificial diet |
| キーワード |
|
|
言語 |
en |
|
主題Scheme |
Other |
|
主題 |
immunoglobulin G |
| 書誌情報 |
en : Biomolecules
巻 10,
号 11,
p. 1482,
発行日 2020-10-26
|
| 収録物ID |
|
|
収録物識別子タイプ |
ISSN |
|
収録物識別子 |
2218273X |
| 出版者 |
|
|
出版者 |
MDPI |
|
言語 |
en |
| 権利情報 |
|
|
言語 |
en |
|
権利情報 |
This article is an open access article distributed under the terms and conditions of the Creative Commons Attribution (CC BY) license (http://creativecommons.org/licenses/by/4.0/). |
| EID |
|
|
識別子 |
373149 |
|
識別子タイプ |
URI |
| 言語 |
|
|
言語 |
eng |
biom_10_11_1482.pdf (2.32 MB)
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