Recombinant production of a functional SARS-CoV-2 spike receptor binding domain in the green algae Chlamydomonas reinhardtii.
Recombinant production of a functional SARS-CoV-2 spike receptor binding domain in the green algae Chlamydomonas reinhardtii.
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DOI：
10.1371/journal.pone.0257089
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发表时间：
2021
期刊：
影响因子：
3.7
通讯作者：
Mayfield SP
中科院分区：
文献类型：
--
作者：
Berndt AJ;Smalley TN;Ren B;Simkovsky R;Badary A;Sproles AE;Fields FJ;Torres-Tiji Y;Heredia V;Mayfield SP
重组病毒蛋白可用于生产疫苗抗原或试剂，以识别患者血清中的抗体。最低限度，这些蛋白质必须正确折叠并有适当的翻译后修饰。在这里，我们报道了在绿藻衣藻中产生SARS-CoV-2刺突蛋白受体结合域(RBD)。融合到荧光报告蛋白的RBD在靶向ER-高尔基体保留或从细胞中分泌时作为完整蛋白积累，而叶绿体定位版本被截断。ER保留的RBD融合蛋白能够与SARS-CoV-2的宿主靶标-人ACE2受体结合，并被哺乳动物细胞产生的重组RBD特异性地竞争，这表明藻类产生的蛋白经过充分的翻译后修饰，可以作为真正的SARS-CoV-2抗原。由于藻类可以非常廉价地大规模生长，这种重组蛋白可能是其他表达平台的低成本替代品。
Recombinant production of viral proteins can be used to produce vaccine antigens or reagents to identify antibodies in patient serum. Minimally, these proteins must be correctly folded and have appropriate post-translation modifications. Here we report the production of the SARS-CoV-2 spike protein Receptor Binding Domain (RBD) in the green algae Chlamydomonas. RBD fused to a fluorescent reporter protein accumulates as an intact protein when targeted for ER-Golgi retention or secreted from the cell, while a chloroplast localized version is truncated. The ER-retained RBD fusion protein was able to bind the human ACE2 receptor, the host target of SARS-CoV-2, and was specifically out-competed by mammalian cell-produced recombinant RBD, suggesting that the algae produced proteins are sufficiently post-translationally modified to act as authentic SARS-CoV-2 antigens. Because algae can be grown at large scale very inexpensively, this recombinant protein may be a low cost alternative to other expression platforms.
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DOI：
10.1016/s2468-2667(21)00055-4
发表时间：
2021-05
期刊：
The Lancet. Public health
影响因子：
--
作者：
Graham MS;Sudre CH;May A;Antonelli M;Murray B;Varsavsky T;Kläser K;Canas LS;Molteni E;Modat M;Drew DA;Nguyen LH;Polidori L;Selvachandran S;Hu C;Capdevila J;COVID-19 Genomics UK (COG-UK) Consortium;Hammers A;Chan AT;Wolf J;Spector TD;Steves CJ;Ourselin S
通讯作者：
Ourselin S
影响因子：
5.6
作者：
Demurtas OC;Massa S;Illiano E;De Martinis D;Chan PK;Di Bonito P;Franconi R
通讯作者：
Franconi R
影响因子：
64.5
作者：
Dejnirattisai W;Zhou D;Ginn HM;Duyvesteyn HME;Supasa P;Case JB;Zhao Y;Walter TS;Mentzer AJ;Liu C;Wang B;Paesen GC;Slon-Campos J;López-Camacho C;Kafai NM;Bailey AL;Chen RE;Ying B;Thompson C;Bolton J;Fyfe A;Gupta S;Tan TK;Gilbert-Jaramillo J;James W;Knight M;Carroll MW;Skelly D;Dold C;Peng Y;Levin R;Dong T;Pollard AJ;Knight JC;Klenerman P;Temperton N;Hall DR;Williams MA;Paterson NG;Bertram FKR;Siebert CA;Clare DK;Howe A;Radecke J;Song Y;Townsend AR;Huang KA;Fry EE;Mongkolsapaya J;Diamond MS;Ren J;Stuart DI;Screaton GR
通讯作者：
Screaton GR
DOI：
10.1016/j.algal.2019.101523
发表时间：
2019-08-01
影响因子：
5.1
作者：
Fields, Francis J.;Ostrand, Joseph T.;Mayfield, Stephen P.
通讯作者：
Mayfield, Stephen P.
影响因子：
2.4
作者：
HwangBo, Kwon;Son, Su Hyun;Jeong, Won Joong
通讯作者：
Jeong, Won Joong