Max-Planck-Institut für Molekulare Pflanzenphysiologie, Department of Organelle Biology, Biotechnology and Molecular Ecophysiology, D-14476 Potsdam-Golm, Germany.
Institute of Biochemistry of the Romanian Academy, Department of Viral Glycoproteins, 060031 Bucharest, Romania.
Proc Natl Acad Sci U S A. 2024 Jun 11;121(24):e2400145121. doi: 10.1073/pnas.2400145121. Epub 2024 Jun 4.
Microalgae are promising production platforms for the cost-effective production of recombinant proteins. We have recently established that the red alga provides superior transgene expression properties, due to the episomal maintenance of transformation vectors as multicopy plasmids in the nucleus. Here, we have explored the potential of to synthesize complex pharmaceutical proteins to high levels. Testing expression constructs for a candidate subunit vaccine against the hepatitis C virus (HCV), we show that the soluble HCV E2 glycoprotein can be produced in transgenic algal cultures to high levels. The antigen undergoes faithful posttranslational modification by N-glycosylation and is recognized by conformationally selective antibodies, suggesting that it adopts a proper antigenic conformation in the endoplasmic reticulum of red algal cells. We also report the experimental determination of the structure of the N-glycan moiety that is attached to glycosylated proteins in . Finally, we demonstrate the immunogenicity of the HCV antigen produced in red algae when administered by injection as pure protein or by feeding of algal biomass.
微藻是具有成本效益的生产平台，可用于生产重组蛋白。我们最近发现，红藻 由于转化载体作为核内多拷贝质粒以附加体形式存在，因此具有优越的转基因表达特性。在这里，我们探索了 合成复杂药物蛋白的潜力，以期达到高水平。我们测试了针对丙型肝炎病毒 (HCV) 的候选亚单位疫苗的表达构建体，结果表明，可溶性 HCV E2 糖蛋白可在转基因藻类培养物中高水平产生。该抗原通过 N-糖基化进行忠实的翻译后修饰，并被构象选择性抗体识别，表明它在红藻细胞的内质网中采用适当的抗原构象。我们还报告了对与 中糖基化蛋白相连的 N-聚糖部分的结构的实验测定。最后，我们证明了当以纯蛋白形式注射或通过藻类生物量喂养给药时，在红藻中产生的 HCV 抗原具有免疫原性。
