An Effective Way of Producing Fully Assembled Antibody in Transgenic Tobacco Plants by Linking Heavy and Light Chains via a Self-Cleaving 2A Peptide.
An Effective Way of Producing Fully Assembled Antibody in Transgenic Tobacco Plants by Linking Heavy and Light Chains via a Self-Cleaving 2A Peptide.
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DOI：
10.3389/fpls.2018.01379
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发表时间：
2018
影响因子：
5.6
通讯作者：
Xie J
中科院分区：
文献类型：
--
作者：
Lin Y;Hung CY;Bhattacharya C;Nichols S;Rahimuddin H;Kittur FS;Leung T;Xie J
治疗性单抗已发展成为治疗各种疾病的一类重要有效药物。由于抗体分子由两条相同的重链(HC)和两条轻链(LC)组成，每条链由两个不同的基因编码，因此它们在相似水平的表达对于高效组装功能性重组单抗至关重要。尽管基于植物的表达系统已经被测试可以生产完全组装的重组单抗，但在转基因植物中以类似水平协调表达HC和LC仍然是一个挑战。口蹄疫病毒(FMDV)2A肽的编码序列已被成功地用于连接两个或更多基因，这使得翻译的多蛋白能够在各种转基因生物中自我切割成单独的蛋白质。在本研究中，我们探索了F2A在埃博拉病毒单抗(EBOV MAb)生产中的应用。我们发现，与在两个独立的转录单元中表达HC和LC的烟草相比，携带含有由2A连接的HC和LC的转录单元的转基因烟草不仅产生了相似水平的HC和LC，而且获得了更高的完全组装的EBOV mAb产量。纯化的埃博拉病毒单抗与埃博拉表位多肽结合，表观Kd值为90.13~149.2 nM，表明其组装正确，与埃博拉表位多肽有较高的亲和力。据我们所知，这是第一个通过在稳定转化的烟草植株中过表达由HC、LC和2A组成的单一转录单位来产生单抗的报道。
Therapeutic monoclonal antibodies (mAbs) have evolved into an important class of effective medicine in treatment of various diseases. Since the antibody molecule consists of two identical heavy chains (HC) and two light chains (LC), each chain encoded by two different genes, their expressions at similar levels are critical for efficient assembly of functional recombinant mAbs. Although the plant-based expression system has been tested to produce fully assembled recombinant mAbs, coordinately expressing HC and LC at similar levels in a transgenic plant remains a challenge. A sequence coding for a foot-and-mouth disease virus (FMDV) 2A peptide has been successfully used to link two or more genes, which enable the translated polyprotein to be “self-cleaved” into individual protein in various genetically modified organisms. In the present study, we exploited the usage of F2A in Ebola virus monoclonal antibody (EBOV mAb) production. We found that transgenic tobacco plants carrying a transcription unit containing HC and LC linked by 2A not only produced similar levels of HC and LC but also rendered a higher yield of fully assembled EBOV mAb compared to those expressing HC and LC in two independent transcription units. Purified EBOV mAb bound to an Ebola epitope peptide with apparent Kd-values of 90.13–149.2 nM, indicating its proper assembly and high affinity binding to Ebola epitope peptide. To our knowledge, this is the first report showing mAb production by overexpressing a single transcription unit consisting of HC, LC and 2A in stable transformed tobacco plants.
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影响因子：
4.7
作者：
Jez, Jakub;Castilho, Alexandra;Grass, Josephine;Vorauer-Uhl, Karola;Sterovsky, Thomas;Altmann, Friedrich;Steinkellner, Herta
通讯作者：
Steinkellner, Herta
影响因子：
46.9
作者：
Chen, LL;Marmey, P;Fauquet, CM
通讯作者：
Fauquet, CM
影响因子：
3.4
作者：
KUO, SC;LAUFFENBURGER, DA
通讯作者：
LAUFFENBURGER, DA
DOI：
10.3390/ma9120994
发表时间：
2016-12-08
期刊：
Materials (Basel, Switzerland)
影响因子：
--
作者：
Choe W;Durgannavar TA;Chung SJ
通讯作者：
Chung SJ
影响因子：
46.9
作者：
Fang, JM;Qian, JJ;Jooss, K
通讯作者：
Jooss, K