The red alga Porphyridium as a host for molecular farming: Efficient production of immunologically active hepatitis C virus glycoprotein

• 成果类型: Article
• 署名作者: Hammel, Alexander; Cucos, Lia-Maria; Caras, Iuliana; Ionescu, Irina; Tucureanu, Catalin; Tofan, Vlad; Costache, Adriana; Onu, Adrian; Hoepfner, Lara; Hippler, Michael; Neupert, Juliane; Popescu, Costin-Ioan; Stavaru, Crina; Branza-Nichita, Norica; Bock, Ralph
• 署名单位: Max Planck Society; Romanian Academy; Institute of Biochemistry, Bucharest; University of Munster; Okayama University; Norwegian Institute of Bioeconomy Research
• 刊物名称: PROCEEDINGS OF THE NATIONAL ACADEMY OF SCIENCES OF THE UNITED STATES OF AMERICA
• ISSN/ISSBN: 0027-10013
• DOI: 10.1073/pnas.2400145121
• 发表日期: 2024-06-11
• 关键词: chlamydomonas-reinhardtii; neutralizing antibodies; endoplasmic-reticulum; expression; gene; antigen; glycosylation; localization; chloroplast; inhibition

摘要

Microalgae are promising production platforms for the cost- effective production of recombinant proteins. We have recently established that the red alga Porphyridium purpureum 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 Porphyridium 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 Porphyridium. 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.