NON-GLYCOSYLATED TRANSFERRIN EXPRESSED IN MONOCOTS
Disclosed are compositions and methods of making non-glycosylated transferrin protein in transgenic monocot plants.
1 . A pharmaceutical composition comprising:
a fusion protein comprising non-glycosylated transferrin and a therapeutic protein, wherein the fusion protein is expressed in and purified from a monocot seed.
2 . A cell culture media composition comprising:
a fusion protein comprising non-glycosylated transferrin and a therapeutic protein wherein the fusion protein is expressed in and purified from a monocot seed.
3 . A transgenic monocot seed-derived composition comprising:
a fusion protein comprising non-glycosylated transferrin and a therapeutic protein wherein the fusion protein is expressed in and purified from a monocot seed.
4 . A food composition comprising:
a fusion protein comprising non-glycosylated transferrin and a therapeutic protein wherein the fusion protein is expressed in and purified from a monocot seed.
5 . The composition of any of claims 1 - 4 , formulated for injectable delivery.
6 . The composition of any of claims 1 - 4 , formulated for oral delivery.
7 . The composition of any of claims 1 - 4 , wherein the therapeutic protein is insulin or proinsulin.
8 . A transgenic monocot seed-derived composition, selected from the group consisting of a whole-seed food composition, a flour composition, an extract composition and a malt composition, comprising a fusion protein comprising non-glycosylated transferrin and a therapeutic protein.
9 . The seed-derived composition of claim 8 , wherein the fusion protein comprising non-glycosylated transferrin and a therapeutic protein constitutes at least 0.1% of the dry weight of the seed-derived composition.
10 . The composition of claim 8 or claim 9 , wherein the therapeutic protein is insulin or proinsulin.
11 . A serum-free cell culture medium comprising an extract of transgenic monocot seed comprising a seed-expressed fusion protein comprising:
a seed-specific signal sequence capable of targeting a polypeptide linked thereto to a protein storage body;
non-glycosylated transferrin; and
a therapeutic protein.
12 . The serum-free cell culture medium of claim 11 , wherein the therapeutic protein is insulin or proinsulin.
13 . A method of producing a recombinant fusion protein comprising transferrin and a therapeutic protein in monocot plant seeds, comprising the steps of:
(a) transforming a monocot plant cell with a chimeric gene comprising
(i) a seed maturation-specific promoter;
(ii) a first DNA sequence, operably linked to said promoter, said first DNA sequence encoding a signal sequence targeting a polypeptide linked thereto to a protein storage body of a monocot plant seed cell; and
(iii) a second DNA sequence, linked in translation frame with the first DNA sequence, encoding a fusion protein comprising transferrin and a therapeutic protein,
wherein the first DNA sequence and the second DNA sequence together encode a fusion protein comprising the signal sequence, a therapeutic protein and transferrin;
(b) growing the monocot plant from the transformed monocot plant cell for a time sufficient to produce seeds containing the fusion protein; and
(c) harvesting the seeds from the plant.
14 . The method of claim 13 , wherein the first DNA sequence encoding a signal sequence targeting a polypeptide linked thereto to a protein storage body of a monocot plant seed cell encodes a glutelin signal sequence.
15 . The method of claim 13 , wherein the fusion protein constitutes at least 0.01% seed weight of the harvested seeds.
16 . A transformed rice plant produced according to the method of claim 13 .
17 . A transformed rice seed produced according to the method of claim 13 .