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Enhanced HIV SOSIP Envelope yields in plants through transient co-expression of peptidyl-prolyl isomerase B and calreticulin chaperones and ER targeting.
Rosenberg, Yvonne J; Jiang, Xiaoming; Lees, Jonathan P; Urban, Lori A; Mao, Lingjun; Sack, Markus.
  • Rosenberg YJ; PlantVax Corporation, Rockville, MD, USA. yjr@plantvax.com.
  • Jiang X; PlantVax Corporation, Rockville, MD, USA.
  • Lees JP; PlantVax Corporation, Rockville, MD, USA.
  • Urban LA; PlantVax Corporation, Rockville, MD, USA.
  • Mao L; PlantVax Corporation, Rockville, MD, USA.
  • Sack M; Pro-SPR GmbH, Alsdorf, Germany.
Sci Rep ; 12(1): 10027, 2022 06 15.
Article in English | MEDLINE | ID: covidwho-1921705
ABSTRACT
High yield production of recombinant HIV SOSIP envelope (Env) trimers has proven elusive as numerous disulfide bonds, proteolytic cleavage and extensive glycosylation pose high demands on the host cell machinery and stress imposed by accumulation of misfolded proteins may ultimately lead to cellular toxicity. The present study utilized the Nicotiana benthamiana/p19 (N.b./p19) transient plant system to assess co-expression of two ER master regulators and 5 chaperones, crucial in the folding process, to enhance yields of three Env SOSIPs, single chain BG505 SOSIP.664 gp140, CH505TF.6R.SOSIP.664.v4.1 and CH848-10.17-DT9. Phenotypic changes in leaves induced by SOSIP expression were employed to rapidly identify chaperone-assisted improvement in health and expression. Up to 15-fold increases were obtained by co-infiltration of peptidylprolvl isomerase (PPI) and calreticulin (CRT) which were further enhanced by addition of the ER-retrieval KDEL tags to the SOSIP genes; levels depending on individual SOSIP type, day of harvest and chaperone gene dosage. Results are consistent with reducing SOSIP misfolding and cellular stress due to increased exposure to the plant host cell's calnexin/calreticulin network and accelerating the rate-limiting cis-trans isomerization of Xaa-Pro peptide bonds respectively. Plant transient co-expression facilitates rapid identification of host cell factors and will be translatable to other complex glycoproteins and mammalian expression systems.
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Full text: Available Collection: International databases Database: MEDLINE Main subject: HIV Infections / HIV-1 Limits: Animals Language: English Journal: Sci Rep Year: 2022 Document Type: Article Affiliation country: S41598-022-14075-3

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Full text: Available Collection: International databases Database: MEDLINE Main subject: HIV Infections / HIV-1 Limits: Animals Language: English Journal: Sci Rep Year: 2022 Document Type: Article Affiliation country: S41598-022-14075-3