Co‐expression of human calreticulin significantly improves the production of HIV gp140 and other viral glycoproteins in plants
| Autor: | Emmanuel Margolin, George P. Lomonossoff, Nobuyuki Matoba, Ann E. Meyers, Ros Chapman, Anna-Lise Williamson, Daniel Ponndorf, Edward P. Rybicki, Hadrien Peyret, Jason Naude, Michiel T. van Diepen, Yulia Meshcheriakova, Young Jun Oh, Matthew Verbeek |
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| Rok vydání: | 2020 |
| Předmět: |
glycoprotein
folding 0106 biological sciences 0301 basic medicine viruses Nicotiana benthamiana virus Plant Science 01 natural sciences Virus calreticulin 03 medical and health sciences chaperone cleavage co‐expression Furin Research Articles Secretory pathway chemistry.chemical_classification biology Endoplasmic reticulum HIV biology.organism_classification Cell biology 030104 developmental biology chemistry Chaperone (protein) biology.protein Glycoprotein furin Agronomy and Crop Science Calreticulin Research Article 010606 plant biology & botany Biotechnology |
| Zdroj: | Plant Biotechnology Journal |
| ISSN: | 1467-7652 1467-7644 |
| DOI: | 10.1111/pbi.13369 |
| Popis: | Summary Plant molecular farming (PMF) is rapidly gaining traction as a viable alternative to the currently accepted paradigm of producing biologics. While the platform is potentially cheaper and more scalable than conventional manufacturing systems, expression yields and appropriate post‐translational modifications along the plant secretory pathway remain a challenge for certain proteins. Viral fusion glycoproteins in particular are often expressed at low yields in plants and, in some cases, may not be appropriately processed. Recently, however, transiently or stably engineering the host plant has shown promise as a strategy for producing heterologous proteins with more complex maturation requirements. In this study we investigated the co‐expression of a suite of human chaperones to improve the production of a human immunodeficiency virus (HIV) type 1 soluble gp140 vaccine candidate in Nicotiana benthamiana plants. The co‐expression of calreticulin (CRT) resulted in a dramatic increase in Env expression and ameliorated the endoplasmic reticulum (ER) stress response ‐ as evidenced by lower transcript abundance of representative stress‐responsive genes. The co‐expression of CRT similarly improved accumulation of glycoproteins from Epstein‐Barr virus (EBV), Rift Valley fever virus (RVFV) and chikungunya virus (CHIKV), suggesting that the endogenous chaperone machinery may impose a bottleneck for their production. We subsequently successfully combined the co‐expression of human CRT with the transient expression of human furin, to enable the production of an appropriately cleaved HIV gp140 antigen. These transient plant host engineering strategies are a promising approach for the production of high yields of appropriately processed and cleaved viral glycoproteins. |
| Databáze: | OpenAIRE |
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