Intracellular reprogramming of expression, glycosylation, and function of a plant-derived antiviral therapeutic monoclonal antibody.
Intracellular reprogramming of expression, glycosylation, and function of a plant-derived antiviral therapeutic monoclonal antibody.
复制标题
DOI：
10.1371/journal.pone.0068772
复制
发表时间：
2013
期刊：
影响因子：
3.7
通讯作者：
Ko K
中科院分区：
文献类型：
--
作者：
Lee JH;Park DY;Lee KJ;Kim YK;So YK;Ryu JS;Oh SH;Han YS;Ko K;Choo YK;Park SJ;Brodzik R;Lee KK;Oh DB;Hwang KA;Koprowski H;Lee YS;Ko K
植物基因工程，这导致了生产植物源性单克隆抗体（mAbPs），提供了一个安全和经济有效的替代传统的抗体表达方法。在这项研究中，抗狂犬病病毒的mAbP SO 57的表达水平和生物学特性进行了分析，有或没有内质网（ER）滞留肽信号（Lys-Asp-Glu-Leu; KDEL）在转基因烟草植物（烟草）。无论转录水平如何，具有KDEL的mAbP SO 57（mAbPK）的表达水平显著高于不具有KDEL的mAbP SO 57（mAbP）的表达水平。纯化mAbP和mAbPK以及杂交瘤衍生mAb（mAbH）的Fc结构域与FcγRI受体（CD 64）的结合活性水平相似。mAbPK具有寡聚甘露糖（OM）型（91.7%）和高尔基型（8.3%）的聚糖谱，而mAbP主要具有高尔基型聚糖（96.8%），与mAbH相似。共聚焦分析显示，mAbPK共定位于ER-示踪信号和细胞核周围的细胞区域，表明mAbP与KDEL在ER中的积累。在BALB/c小鼠中，mAbP和mAbPK均以与mAbH相似的趋势消失。此外，mAbPK在中和狂犬病病毒CVS-11的活性方面与mAbH一样有效。这些结果表明，重组mAbP通过KDEL的ER定位重新编程OM糖基化，并增强植物中功能性抗病毒治疗性抗体的产生。
Plant genetic engineering, which has led to the production of plant-derived monoclonal antibodies (mAbPs), provides a safe and economically effective alternative to conventional antibody expression methods. In this study, the expression levels and biological properties of the anti-rabies virus mAbP SO57 with or without an endoplasmic reticulum (ER)-retention peptide signal (Lys-Asp-Glu-Leu; KDEL) in transgenic tobacco plants (Nicotiana tabacum) were analyzed. The expression levels of mAbP SO57 with KDEL (mAbPK) were significantly higher than those of mAbP SO57 without KDEL (mAbP) regardless of the transcription level. The Fc domains of both purified mAbP and mAbPK and hybridoma-derived mAb (mAbH) had similar levels of binding activity to the FcγRI receptor (CD64). The mAbPK had glycan profiles of both oligomannose (OM) type (91.7%) and Golgi type (8.3%), whereas the mAbP had mainly Golgi type glycans (96.8%) similar to those seen with mAbH. Confocal analysis showed that the mAbPK was co-localized to ER-tracker signal and cellular areas surrounding the nucleus indicating accumulation of the mAbP with KDEL in the ER. Both mAbP and mAbPK disappeared with similar trends to mAbH in BALB/c mice. In addition, mAbPK was as effective as mAbH at neutralizing the activity of the rabies virus CVS-11. These results suggest that the ER localization of the recombinant mAbP by KDEL reprograms OM glycosylation and enhances the production of the functional antivirus therapeutic antibody in the plant.
登录
查看更多内容
DOI：
10.1007/s004380050932
发表时间：
1999-01-01
期刊：
MOLECULAR AND GENERAL GENETICS
影响因子：
--
作者：
De Neve, M;De Buck, S;Depicker, A
通讯作者：
Depicker, A
影响因子：
5.5
作者：
Cliquet, F.;Gurbuxani, J. P.;Barrat, J.
通讯作者：
Barrat, J.
DOI：
10.1016/j.bbrc.2009.01.070
发表时间：
2009-03-06
影响因子：
3.1
作者：
Lee, Kyung Jin;Jung, Jin-Hee;Oh, Doo-Byoung
通讯作者：
Oh, Doo-Byoung
影响因子：
5.8
作者：
Hu, Rongliang;Zhang, Shoufeng;Xiao, Yueqiang
通讯作者：
Xiao, Yueqiang
影响因子：
4.3
作者：
GarciaCasado, G;SanchezMonge, R;Gomez, L
通讯作者：
Gomez, L