Productions of hetero-oligomeric proteins in plants

Abstract

Process of producing in a plant, in plant tissue, or in plant cells a hetero-oligomeric protein comprising at least a first and a second protein subunit, said process comprising expressing in plant cells at least said first and said second protein subunit by (i) providing to said plant, said plant tissue or said plant cells a plus-sense single-stranded RNA viral vector encoding at least said first and said second protein subunit or (ii) providing to said plant, said plant tissue or said plant cells a first and a second plus-sense single-stranded RNA viral vector, said first viral vector encoding at least said first protein subunit, said second viral vector encoding at least said second protein subunit, whereby at least said first viral vector and said second viral vector are non-competing viral vectors.

1 - 42 . (canceled) 
     
     
         43 . A process of producing in a plant, in plant tissue, or in plant cells a hetero-oligomeric protein comprising at least a first and a second protein subunit, said process comprising expressing in plant cells at least said first and said second protein subunit by providing to said plant, said plant tissue or said plant cells a first and a second plus-sense single-stranded RNA viral vector, said first viral vector encoding at least said first protein subunit, said second viral vector encoding at least said second protein subunit, whereby said first viral vector and said second viral vector are
 (a) non-competing viral vectors;   (b) non-competing viral vectors in that they are not derived from viruses of the same virus species;   (c) non-competing viral vectors in that they differ in sequence portions other than sequence portions coding for said first and said second protein subunits;   (d) non-competing viral vectors in that the replicase ORF(s) of said first viral vector and the replicase ORF(s) of said second viral vector have a homology of at most 90%; or   (e) non-competing viral vectors in that the replicase ORF(s) of said first viral vector and the replicase ORF(s) of said second viral vector have an identity of at most 85%.   
     
     
         44 . The process according to  claim 43 , followed by isolating said hetero-oligomeric protein from said plant, said plant tissue, or said plant cells. 
     
     
         45 . The process according to  claim 43 , wherein said first viral vector contains a first heterologous sequence encoding said first protein subunit and said second viral vector contains a second heterologous sequence encoding said second protein subunit, whereby expression of said first and/or of said second protein subunit is under the control of a sub-genomic promoter. 
     
     
         46 . The process according to  claim 43 , wherein said first viral vector and/or said second viral vector is/are devoid of a
 functional coat protein ORF,   a functional movement protein ORF, and/or   a functional origin of viral particle assembly.   
     
     
         47 . The process according to  claim 43 , wherein said first viral vector is derived from a virus belonging to the genus Potexvirus and said second viral vector is derived from a virus belonging to the genus Potyvirus. 
     
     
         48 . The process according to  claim 47 , wherein said first viral vector is derived from Potato Virus X and said second viral vector is derived from Potato Virus Y. 
     
     
         49 . The process according to  claim 43 , wherein said first viral vector is derived from a virus belonging to the genus Potexvirus and said second viral vector is derived from a virus belonging to the genus Tobamovirus. 
     
     
         50 . The process according to  claim 49 , wherein said first viral vector is derived from Potato Virus X and said second viral vector is derived from Tobacco Mosaic Virus. 
     
     
         51 . The process according to  claim 43 , wherein said first and said second viral vectors have at most 60% sequence homology. 
     
     
         52 . The process according to  claim 43 , wherein said hetero-oligomeric protein is an antibody. 
     
     
         53 . The process according to  claim 43 , wherein said viral vector(s) is (are) transiently provided to said plant, plant tissue, or plant cells. 
     
     
         54 . The process according to  claim 53 , wherein said viral vector(s) is (are) transiently provided to said plant, plant tissue, or plant cells by  Agrobacterium  transfection. 
     
     
         55 . The process according to  claim 43 , wherein said RNA viral vector(s) is (are) stably incorporated into plant chromosomal DNA as DNA precursors of said RNA viral vectors. 
     
     
         56 . The process according to  claim 55 , wherein controlled release of said RNA viral vector(s) from said DNA precursors is (are) provided by inducible promoters. 
     
     
         57 . The process according to  claim 52 , wherein said antibody is an immunoglobulin comprising at least a portion of an antigen binding domain. 
     
     
         58 . The method according to  claim 52 , wherein said antibody or its derivative belongs to the immunoglobulin G class, to the immunoglobulin A class, to the immunoglobulin M class, to the immunoglobulin D class, or to the immunoglobulin E class. 
     
     
         59 . The process according to  claim 43 , wherein said hetero-oligomeric protein is insulin. 
     
     
         60 . The process according to  claim 43 , wherein at least one or at least two or more subunits of said hetero-oligomeric protein contain(s) an endoplasmatic reticulum retention signal KDEL. 
     
     
         61 . The process according to  claim 43 , wherein said plant is a monocot or a dicot plant. 
     
     
         62 . The process according to  claim 61 , wherein said dicot plant belongs to the Solanacea family, Brassicacea family, the Legume family, or to the family Chenopodiaceae.