Institute of Molecular and Cellular Plant Biology (IBMCP), Spanish Research Council Agency (CSIC), Polytechnic University of Valencia (UPV), Avda Tarongers SN, Valencia 46022, Spain.
Int J Mol Sci. 2013 Mar 18;14(3):6205-22. doi: 10.3390/ijms14036205.
Delivery of secretory immunoglobulin A (sIgA) to mucosal surfaces as a passive immunotherapy agent is a promising strategy to prevent infectious diseases. Recombinant sIgA production in plants requires the co-expression of four transcriptional units encoding the light chain (LC), heavy chain (HC), joining chain (JC) and secretory component (SC). As a way to optimize sIgA production in plants, we tested the combinatorial expression of 16 versions of a human sIgA against the VP8* rotavirus antigen in Nicotiana benthamiana, using the recently developed GoldenBraid multigene assembly system. Each sIgA version was obtained by combining one of the two types of HC (α1 and α2) with one of the two LC types (k and λ) and linking or not a KDEL peptide to the HC and/or SC. From the analysis of the anti-VP8* activity, it was concluded that those sIgA versions carrying HCα1 and LCλ provided the highest yields. Moreover, ER retention significantly increased antibody production, particularly when the KDEL signal was linked to the SC. Maximum expression levels of 32.5 μg IgA/g fresh weight (FW) were obtained in the best performing combination, with an estimated 33% of it in the form of a secretory complex.
将分泌型免疫球蛋白 A（sIgA）递送到黏膜表面作为一种被动免疫疗法是预防传染病的一种很有前途的策略。在植物中生产重组 sIgA 需要共表达四个转录单元，分别编码轻链（LC）、重链（HC）、连接链（JC）和分泌成分（SC）。为了优化植物中 sIgA 的生产，我们使用最近开发的 GoldenBraid 多基因组装系统，在 Nicotiana benthamiana 中测试了针对 VP8轮状病毒抗原的 16 种人 sIgA 组合表达。每种 sIgA 版本都是通过将两种 HC 类型（α1 和 α2）之一与两种 LC 类型（k 和 λ）之一组合，并将 KDEL 肽连接或不连接到 HC 和/或 SC 上来获得的。从抗 VP8活性分析的结果来看，携带 HCα1 和 LCλ的 sIgA 版本提供了最高的产量。此外，内质网保留显著增加了抗体的产生，尤其是当 KDEL 信号与 SC 连接时。在表现最好的组合中，获得了 32.5 μg IgA/g 鲜重（FW）的最高表达水平，其中约 33%以分泌复合物的形式存在。
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