Flexible approaches are required for successful production of recombinant proteins in plants

Flexible approaches are required for successful production of recombinant proteins in plants

Production of recombinant proteins has increased significantly due to the ever-increasing demand in research laboratories, medical fields, and agriculture. Despite a variety of recombinant protein expression systems and gene engineering procedures exist owever, there is a limitation in achieving sufficient quantities of active, properly folded recombinant proteins. The selection of an appropriate expression system for the production of recombinant proteins should generally be chosen based on the biochemical and biological properties of the desired proteins to be produced. Plant expression platforms offer commercially inexpensive, safe, and fast scale-up properties for the production of recombinant proteins compared to traditional mammalian cell expression systems. However, the absence of some mammalian posttranslational modifications (PTMs) and aberrant N-glycosylation limit the use of these production systems. Therefore, molecular engineering of plant secretory pathways has provided an opportunity to develop next-generation pharmaceuticals with targeted biological functions. In this mini-review, we discuss the approaches that are needed for the production of functional recombinant proteins such as vaccines, antibodies, enzymes for medical, agricultural, and industrial applications.

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  • 1. Nandi S, Kwong AT, Holtz BR, et al. Techno-economic analysis of a transient plant-based platform for monoclonal antibody production. mAbs. 2016;8:1456-66.
  • 2. Gomord V, Faye L. Posttranslational modification of therapeutic proteins in plants. Curr Opin Plant Biol. 2004;7:171-81.
  • 3. Chichester JA, Manceva SD, Rhee A, et al. A plant-produced protective antigen vaccine confers protection in rabbits against a lethal aerosolized challenge with Bacillus anthracis Ames spores. Hum Vaccin Immunother. 2013;9:544–52.
  • 4. Mamedov T, Ghosh A, Jones RM, et al. Production of non-glycosylated recombinant proteins in Nicotiana benthamiana plants by co-expressing bacterial PNGase F. Plant Biotechnol J. 2012;10:773-82.
  • 5. Mamedov T, Yusibov V. In vivo deglycosylation of recombinant proteins in plants by co-expression with bacterial PNGase F. Bioengineered. 2013;4:338-42.
  • 6. Mamedov T, Chichester JA, Jones RM, et al. Production of functionally active and immunogenic non-glycosylated protective antigen from bacillus anthracis in nicotiana benthamiana by co-expression with peptide-nglycosidase f (PNGase F) of flavobacterium meningosepticum. PLoS One. 2016;11:e0153956.
  • 7. Fischer R, Buyel JF. Molecular farming - The slope of enlightenment. Biotechnol Adv. 2020;40:107519. 8. Castilho A, Strasser R, Stadlmann J, et al. In planta protein sialylation through overexpression of the respective mammalian pathway. J Chem Biol. 2010;285:15923-930.
  • 9. Grosse-Holz F, Madeira L, Zahid MA, et al. Three unrelated protease inhibitors enhance accumulation of pharmaceutical recombinant proteins in Nicotiana benthamiana. Plant Biotechnol J. 2018;16:1797–810.
  • 10. Gomord V, Sourrouille C, Fitchette AC, et al. Production and glycosylation of plant-made pharmaceuticals: the antibodies as a challenge. Plant Biotechnol J. 2004;2:83-100.
  • 11. Gomord V, Denmat LA, Fitchette-Lainé AC, et al. The C-terminal HDEL sequence is sufficient for retention of secretory proteins in the endoplasmic reticulum (ER) but promotes vacuolar targeting of proteins that escape the ER. Plant J. 1997;11:313-25.
  • 12. Saint-Jore-Dupas C, Faye L, Gomord V. From planta to pharma with glycosylation in the toolbox. Trends Biotechnol. 2007;25:317-23.
  • 13. Strasser R, Castilho A, Stadlmann J, et al. Improved virus neutralization by plant-produced anti-HIV antibodies with a homogeneous beta1,4- galactosylated N-glycan profile. J Biol Chem. 2009;284:20479–485.
  • 14. Lerouge P, Cabanes-Macheteau M, Rayon C, et al. N-glycoprotein biosynthesis in plants: recent developments and future trends. Plant Mol Biol. 1998;38:31-48.
  • 15. Pagny S, Cabanes-Macheteau M, Gillikin JW, et al. Protein recycling from the Golgi apparatus to the endoplasmic reticulum in plants and its minor contribution to calreticulin retention. Plant Cell. 2000;12:739-56.
  • 16. Sriraman R, Bardor M, Sack M, et al. Recombinant anti-hCG antibodies retained in the endoplasmic reticulum of transformed plants lack core-xylose and core-alpha(1,3)-fucose residues. Plant Biotechnol J. 2004;2:279-87.
  • 17. Zeng Y, He X, Danyukova T, et al. Toward Engineering the Mannose 6-Phosphate Elaboration Pathway in Plants for Enzyme Replacement Therapy of Lysosomal Storage Disorders. J Clin Med. 2019;812:2190.
  • 18. Shaaltiel Y, Bartfeld D, Hashmueli S, et al. Production of glucocerebrosidase with terminal mannose glycans for enzyme replacement therapy of Gaucher's disease using a plant cell system. Plant Biotechnol J. 2007;5:579-90.
  • 19. Van Patten SM, Hughes H, Huff MR, et al. Effect of mannose chain length on targeting of glucocerebrosidase for enzyme replacement therapy of Gaucher disease. Glycobiology. 2007;17:467-78.
  • 20. Gomord V, Fitchette AC, Menu-Bouaouiche L, et al. Plant-specific glycosylation patterns in the context of therapeutic protein production. Plant Biotechnol J. 2010;8:564-87.
  • 21. Helenius A, Aebi M. Intracellular functions of N-linked glycans. Science. 2001;291:2364-9.
  • 22. Helenius A, Aebi M. Roles of N-linked glycans in the endoplasmic reticulum. Annu Rev Biochem. 2004;73:1019-49.
  • 23. Wilson IB, Zeleny R, Kolarich D, et al. Analysis of Asn-linked glycans from vegetable foodstuffs: widespread occurrence of Lewis a, core alpha1,3- linked fucose and xylose substitutions. Glycobiology. 2001;11:261-74.
  • 24. Li J, Stoddard TJ, Demorest ZL, et al. Multiplexed, targeted gene editing in Nicotiana benthamiana for glyco-engineering and monoclonal antibody production. Plant Biotechnol J. 2016;14:533-42.
  • 25. Hanania U, Ariel T, Tekoah Y, Fux L, et al. Establishment of a tobacco BY2 cell line devoid of plant-specific xylose and fucose as a platform for the production of biotherapeutic proteins. Plant Biotechnol J. 2017;15:1120-29.
  • 26. Koprivova A, Stemmer C, Altmann F, Hoffmann A. Targeted knockouts of Physcomitrella lacking plant-specific immunogenic N-glycans. Plant Biotechnol J. 2004;2:517-23.
  • 27. Cox KM, Sterling JD, Regan JT, Gasdaska JR, et al. Glycan optimization of a human monoclonal antibody in the aquatic plant Lemna minor. Nat. Biotechnol. 2006;24:1591–97.
  • 28. Jansing J, Sack,M, Augustine SM, Fischer R, Bortesi L. CRISPR/ Cas9-mediated knockout of six glycosyltransferase genes in Nicotiana benthamiana for the production of recombinant proteins lacking β-1,2- xylose and core α-1,3-fucose. Plant Biotechnol J. 2020;17:350-61.
  • 29. Junttila TT, Parsons K, Olsson C, et al. Superior in vivo efficacy of afucosylated trastuzumab in the treatment of HER2-amplified breast cancer. Cancer Res. 2010;70:4481-9.
  • 30. Gasdaska JR, Sherwood S, Regan JT, Dickey LF. An afucosylated anti-CD20 monoclonal antibody with greater antibody-dependent cellular cytotoxicity and B-cell depletion and lower complement-dependent cytotoxicity than rituximab. Mol Immunol. 2012;50:134-41.
  • 31. Salles G, Morschhauser F, Lamy T, et al. Phase 1 study results of the type II glycoengineered humanized anti-CD20 monoclonal antibody obinutuzumab (GA101) in B-cell lymphoma patients. Blood.2012;119:5126-32.
  • 32. Liebminger E, Veit C, Pabst M, et al. Beta-N-acetylhexosaminidases HEXO1 and HEXO3 are responsible for the formation of paucimannosidic N-glycans in Arabidopsis thaliana. J Biol Chem. 2011;286:10793-802.
  • 33. Melo NS, Nimtz M, Conradt HS, et al. Identification of the human Lewis(a) carbohydrate motif in a secretory peroxidase from a plant cell suspension culture (Vaccinium myrtillus L.). FEBS Lett. 1997;41:186–91.
  • 34. Yang WH, Aziz PV, Heithoff DM, et al. An intrinsic mechanism of secreted protein aging and turnover. Proc Natl Acad Sci U S A. 2015;112:13657-62.
  • 35. Shin YJ, Castilho A, Dicker M, et al. Reduced paucimannosidic N-glycan formation by suppression of a specific β-hexosaminidase from Nicotiana benthamiana. Plant Biotechnol J. 2017;15:197-206.
  • 36. Fitchette-Lainé AC, Gomord V, Cabanes M, et al. N-glycans harboring the Lewis a epitope are expressed at the surface of plant cells. Plant. 1997;12:1411-7.
  • 37. Strasser R, Stadlmann J, Svoboda B, et al. Molecular basis of N-acetylglucosaminyltransferase I deficiency in Arabidopsis thaliana plants lacking complex N-glycans. Biochem J. 2005;387:385-91.
  • 38. Parsons J, Altmann F, Arrenberg CK, et al. Moss-based production of asialoerythropoietin devoid of Lewis A and other plant-typical carbohydrate determinants. Plant Biotechnol J. 2012;10:851-61.
  • 39. Meuris L, Santens F, Elson G, Festjens N, et al. GlycoDelete engineering of mammalian cells simplifies N-glycosylation of recombinant proteins. Nat Biotechnol. 2004;32:485-9.
  • 40. Piron R, Santens F, De Paepe A, et al. Using GlycoDelete to produce proteins lacking plant-specific N-glycan modification in seeds. Nat Biotechnol. 2015;33:1135-7.
  • 41. Matsui T, Takita E, Sato T, et al. N-glycosylation at noncanonical Asn-XCys sequences in plant cells. Glycobiology. 2001;21:994-9.
  • 42. Zielinska DF, Gnad F, Wiśniewski JR, Mann M. Precision mapping of an in vivo N-glycoproteome reveals rigid topological and sequence constraints. Cell. 2010;141:897-907.
  • 43. Bakker H, Bardor M, Molthoff JW, Gomord V, et al. Galactoseextended glycans of antibodies produced by transgenic plants. Proc Natl Acad Sci U S A. 2001;98:2899-904.
  • 44. Montero-Morales L, Steinkellner H. Advanced plant-based glycan engineering. Front Bioeng Biotechnol. 2018;6:81.
  • 45. Kallolimath S, Castilho A, Strasser R, et al. Engineering of complex protein sialylation in plants. Proc Natl Acad Sci U S A. 2016;113:9498-503. 46. Sato C, Kitajima K. Disialic, oligosialic and polysialic acids: distribu tion, functions and related disease. J Biochem. 2013;154:115-36.
  • 47. Castilho A, Neumann L, Daskalova S, et al. Engineering of sialylated mucin-type O-glycosylation in plants. J Biol Chem. 2012;287:36518-26.
  • 48. Schjoldager KT, Clausen H. Site-specific protein O-glycosylation modulates proprotein processing - deciphering specific functions of the large polypeptide GalNAc-transferase gene family. Biochim Biophys Acta. 2012;1820:2079-94.
  • 49. Parsons J, Altmann F, Graf M, Stadlmann J. A gene responsible for prolylhydroxylation of moss-produced recombinant human erythropoietin. Sci Rep. 2016;22:3019.
  • 50. Schiavinato M, Strasser R, Mach L, Dohm JC. Genome and transcriptome characterization of the glycoengineered Nicotiana benthamiana line ΔXT/ FT. BMC Genom Data. 2019;20:594.
  • 51. Rouwendal GJ, Wuhrer M, Florack DE, et al. Efficient introduction of a bisecting GlcNAc residue in tobacco N-glycans by expression of the gene encoding human N-acetylglucosaminyltransferase III. Glycobiology. 2007;17:334-44.
  • 52. Wang J, Zhang Y, Wei J, et al. Lewis X oligosaccharides targeting to DC-SIGN enhanced antigen-specific immune response. Immunology. 2007;121:174-82.
  • 53. Mamedov T, Cicek K, Gulec B, et al. In vivo production of non-glycosylated recombinant proteins in Nicotiana benthamiana plants by co-expression with Endo-β-N-acetylglucosaminidase H (Endo H) of Streptomyces plicatus. PLoS One. 2017;12:e0183589.
  • 54. Mamedov T, Musayeva I, Acsora R, et al. Engineering, and production of functionally active human Furin in N. benthamiana plant: In vivo posttranslational processing of target proteins by Furin in plants. PLoS One. 2019a;14:e0213438.
  • 55. Mamedov T, Cicek K, Miura K, et al. A Plant-Produced in vivo deglycosylated full-length Pfs48/45 as a Transmission-Blocking Vaccine Candidate against malaria. Sci Rep. 2019b;9:9868.
  • 56. Nakayama K. Furin: a mammalian subtilisin/Kex2p-like endoprotease involved in processing of a wide variety of precursor proteins. Biochem J. 1997;327:625–35.
  • 57. Lee JH, Andrabi R, Su CY, et al. A broadly neutralizing antibody targets the dynamic hiv envelope trimer apex via a long, rigidified, and anionic β-Hairpin structure. Immunity. 2017;46:690-702.
  • 58. Loos A, Gach JS, Hackl T, et al. Glycan modulation and sulfoengineering of anti-HIV-1 monoclonal antibody PG9 in plants. Proc Natl Acad Sci U S A. 2015;112:12675-80.
  • 59. Hildebrandt H, Dityatev A. Polysialic acid in brain development and synaptic plasticity. Top Curr Chem. 2015;366:55-96.
  • 60. Mercx S, Smargiasso N, Chaumont F, De Pauw E, et al. Inactivation of the beta(1,2)-xylosyltransferase and the alpha(1,3)-fucosyltransferase genes in Nicotiana tabacum BY-2 cells by a Multiplex CRISPR/Cas9 strategy results in glycoproteins without plant-specific glycans. Front Plant Sci 2017; 8:403.
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  • ISSN: 2147-0634
  • Yayın Aralığı: Yılda 4 Sayı
  • Başlangıç: 2012
  • Yayıncı: Effect Publishing Agency ( EPA )