Human blood proteins expressed in monocot seeds

Abstract

The invention is directed to blood proteins produced in monocot seeds and isolated therefrom for use in therapeutic compositions, and to methods of making these isolated blood proteins and to therapeutic compositions comprising them.

1 - 6 . (canceled) 
     
     
         7 . A purified human blood protein obtained by producing a recombinant human blood Protein in monocot plant seeds, comprising the steps of:
 (a) transforming a monocot plant cell with a chimeric gene comprising
 (i) a promoter from the gene of a maturation-specific monocot plant storage protein, 
 (ii) a first DNA sequence, operably linked to said promoter, encoding a monocot plant seed-specific signal sequence capable of targeting a polypeptide linked thereto to a monocot plant seed endosperm cell, and 
 (iii) a second DNA sequence, linked in translation frame with the first DNA sequence, encoding a human blood protein, wherein the first DNA sequence and the second DNA sequence together encode a fusion protein comprising an N-terminal signal sequence and the human blood-Protein: 
   (b) growing a monocot plant from the transformed monocot plant cell for a time sufficient to Produce seeds containing the human blood Protein: and   (c) harvesting the seeds from the plant, wherein the human blood protein constitutes at least about 3.0% of the total soluble protein in the harvested seeds, and   (d) purifying the human blood protein from the harvested seeds,   wherein the human blood protein comprises one or more plant glycosyl groups.   
     
     
         8 . The human blood protein of  claim 7 , selected from the group consisting of hemoglobin, alpha-1-antitrypsin, fibrinogen, human serum albumin, thrombin, antithrombin III, an antibody, and a blood coagulation factor. 
     
     
         9 - 12 . (canceled) 
     
     
         13 . A composition comprising a purified human blood protein comprising at least one plant glycosyl group and at least one pharmaceutically acceptable excipient or nutrient, wherein the human blood protein is produced in a monocot plant containing a nucleic acid sequence encoding the human blood protein and is purified from seed harvested from the monocot plant, and wherein the at least one nutrient is from a source other than the monocot plant. 
     
     
         14 . The composition of  claim 13 , wherein the human blood protein is selected from the group consisting of hemoglobin, alpha-1-antitrypsin, fibrinogen, human serum albumin, thrombin, antithrombin III, an antibody, and a blood coagulation factor. 
     
     
         15 . The composition of  claim 13 , wherein the composition is substantially free of contaminants of the monocot plant. 
     
     
         16 . The composition of  claim 13 , wherein the at least one nutrient is selected from the group consisting of salts, saccharides, vitamins, minerals, amino acids, peptides, and proteins other than the human blood protein. 
     
     
         17 . The composition of  claim 13 , wherein the composition is formulated for parenteral, enteric, inhalation, intranasal or topical delivery. 
     
     
         18 . The composition of  claim 13 , wherein the composition is formulated for enteric delivery, contains at least one pharmaceutically acceptable excipient, and is non-nutritional. 
     
     
         19 . An antibody obtained by producing a recombinant antibody in monocot plant seeds, comprising the steps of:
 (a) transforming a monocot plant cell with a chimeric gene comprising
 (i) a promoter from the gene of a maturation-specific monocot plant storage protein, 
 (ii) a first DNA sequence, operably linked to said promoter, encoding a monocot plant seed-specific signal sequence capable of targeting a polypeptide linked thereto to a monocot plant seed endosperm cell, and 
 (iii) a second DNA sequence, linked in translation frame with the first DNA sequence, encoding an antibody, wherein the first DNA sequence and the second DNA sequence together encode a fusion protein comprising an N-terminal signal sequence and the antibody; 
   (b) growing a monocot plant from the transformed monocot plant cell for a time sufficient to produce seeds containing the antibody; and   (c) harvesting the seeds from the plant,   wherein the antibody comprises one or more plant glycosyl groups.   
     
     
         20 . The antibody of  claim 19 , wherein the antibody is selected from the group consisting of IgG, IgA, and IgM. 
     
     
         21 . The antibody of  claim 19 , wherein the antibody constitutes at least about 5.0% of the total soluble protein in the harvested seeds. 
     
     
         22 . The antibody of  claim 19 , wherein the antibody constitutes at least about 10.0% of the total soluble protein in the harvested seeds. 
     
     
         23 . The antibody of  claim 19 , further comprising purifying the antibody from the harvested seeds. 
     
     
         24 . An antibody obtained by producing a recombinant antibody in monocot plant seeds, comprising the steps of:
 (a) transforming a monocot plant cell with a first expression construct and a second expression construct, each comprising a chimeric gene which is different for each construct, each chimeric gene comprising
 (i) a promoter from the gene of a maturation-specific monocot plant storage protein, 
 (ii) a first DNA sequence, operably linked to said promoter, encoding a monocot plant seed-specific signal sequence capable of targeting a polypeptide linked thereto to a monocot plant seed endosperm cell, and 
 (iii) a second DNA sequence, linked in translation frame with the first DNA sequence, encoding a portion of an antibody, wherein the first DNA sequence and the second DNA sequence together encode a fusion protein comprising an N-terminal signal sequence and the portion of the antibody, wherein the portions of the antibody are assembled to form the antibody; 
   (b) growing a monocot plant from the transformed monocot plant cell for a time sufficient to produce seeds containing the antibody; and   (c) harvesting the seeds from the plant,   wherein the antibody comprises one or more plant glycosyl groups.   
     
     
         25 . The antibody of  claim 24 , wherein the antibody is selected from the group consisting of IgG, IgA, and IgM. 
     
     
         26 . The antibody of  claim 24 , wherein the antibody constitutes at least about 5.0% of the total soluble protein in the harvested seeds. 
     
     
         27 . The antibody of  claim 24 , wherein the antibody constitutes at least about 10.0% of the total soluble protein in the harvested seeds. 
     
     
         28 . The antibody of  claim 24 , further comprising purifying the antibody from the harvested seeds.