PLANT-BASED SYNTHESIS PRODUCTS
The present disclosure provides composition, systems, devices and methods for plant-based production of non-plant proteins through the use of heterologous genes for the expression of said non-plant proteins. Non-plant proteins can include, but are not limited to, mammalian proteins, cytokines, or growth factors.
1 - 77 . (canceled)
78 . A plant cell, wherein the plant cell comprises:
a vector comprising:
(i) a polynucleotide sequence encoding for a heterologous protease inhibitor or a functional variant thereof; and
(ii) a polynucleotide sequence encoding for a mammalian protein or a functional variant thereof,
wherein the mammalian protein is a mammalian growth factor protein selected from the group consisting of mammalian fibroblast growth factor 2 (FGF-2), mammalian insulin-like growth factor 1 (IGF-1), and mammalian vascular endothelial growth factor (VEGF).
79 . The plant cell of claim 78 , wherein the heterologous protease inhibitor is SICYS8.
80 . The plant cell of claim 78 , wherein the vector is:
(i) a viral vector; or
(ii) a bacterial vector.
81 . The plant cell of claim 78 , wherein the vector is Tobacco mosaic virus.
82 . The plant cell of claim 78 , wherein the vector is an Agrobacterium species.
83 . The plant cell of claim 78 , wherein the plant cell is a tobacco plant cell.
84 . The plant cell of claim 83 , wherein the tobacco plant cell is an Nicotiana benthamiana cell.
85 . The plant cell of claim 78 , wherein the mammalian protein is the mammalian FGF-2, and wherein the mammalian FGF-2 is human FGF-2 or bovine FGF-2.
86 . The plant cell of claim 85 , wherein the polynucleotide sequence encoding for the human FGF-2 comprises a sequence with at least 75% sequence identity to SEQ ID NOs: 23, 24, 25, 26, or 28.
87 . The plant cell of claim 85 , wherein the human FGF-2 comprises a polypeptide sequence selected from the group consisting of SEQ ID NOs: 1, 2, 3, 4, 5, 6, and 14.
88 . The plant cell of claim 85 , wherein the bovine FGF-2 comprises a polypeptide sequence of SEQ ID NO: 15.
89 . The plant cell of claim 78 , wherein the mammalian protein is the mammalian IGF-1.
90 . The plant cell of claim 12 , wherein the mammalian IGF-1 comprises a polypeptide sequence of SEQ ID NO: 12 or SEQ ID NO: 13.
91 . The plant cell of claim 78 , wherein the mammalian protein is the mammalian VEGF.
92 . The plant cell of claim 78 , wherein the mammalian VEGF comprises a polypeptide sequence of SEQ ID NO: 20.
93 . A method of manufacturing a mammalian protein, comprising:
(i) culturing a plurality of plant cells, each of the plurality of plant cells comprising a vector comprising:
(a) a polynucleotide sequence encoding for a heterologous protease inhibitor or a functional variant thereof, and
(b) a polynucleotide sequence encoding for a mammalian protein or a functional variant thereof; and
(ii) extracting soluble proteins from a harvested plurality of plant cells, thereby generating an extraction product comprising the mammalian protein or a functional variant thereof,
wherein the mammalian protein is a mammalian growth factor protein selected from the group consisting of mammalian fibroblast growth factor 2 (FGF-2), mammalian insulin-like growth factor 1 (IGF-1), and mammalian vascular endothelial growth factor (VEGF).
94 . The method of claim 93 , further comprising: prior to and/or during (i), infiltrating the vector into another plurality of plant cells, thereby forming the plurality of plant cells.
95 . The method of claim 94 , wherein the infiltrating is vacuum infiltrating.
96 . The method of claim 93 , wherein the mammalian protein is human FGF-2, bovine FGF-2, bovine IGF-1, or human VEGF.
97 . The method of claim 93 , wherein the mammalian protein is histidine-tagged, and wherein the extracting comprises using metal-based histidine-tag affinity (NTA) magnetic beads.