Unstructured recombinant polymers and uses thereof
View Patent ↗The present invention provides unstructured recombinant polymers (URPs) and proteins containing one or more of the URPs. The present invention also provides microproteins, toxins and other related proteinaceous entities, as well as genetic packages displaying these entities. The present invention also provides recombinant polypeptides including vectors encoding the subject proteinaceous entities, as well as host cells comprising the vectors. The subject compositions have a variety of utilities including a range of pharmaceutical applications.
1. An isolated unstructured recombinant polypeptide (URP) comprising at least 40 contiguous amino acids, wherein
(a) the sum of glycine (G), aspartate (D), alanine (A), serine (S), threonine (T), glutamate (E) and proline (P) residues contained in the URP, constitutes at least 80% of the total amino acids of the URP, and the remainder, when present, consists of arginine or lysine, and the remainder does not contain methionine, cysteine, asparagine, and glutamine;
(b) wherein said URP comprises at least three different types of amino acids;
(c) at least 50% of the at least 40 contiguous amino acids in said URP are devoid of secondary structure as determined by Chou-Fasman algorithm;
(d) wherein the URP has a Tepitope score less than −4; and wherein
incorporation of the URP into a heterologous protein results in a fusion protein having at least a 2-fold increase in apparent molecular weight of the fusion protein as compared to the protein lacking said URP, wherein the apparent molecular weight is approximated by size exclusion chromatography, and wherein the fusion protein exhibits an increased half-life as compared to the protein lacking said URP.
2. The unstructured recombinant polypeptide (URP) of claim 1 , wherein said URP has an in vitro serum degradation half-life greater than about 24 hours.
3. The URP of claim 1 , wherein the URP is selected for incorporation into an N- and/or C-terminus of a heterologous protein to create a monomeric fusion protein, and wherein upon incorporation the URP into a heterologous protein, said heterologous protein exhibits a longer serum half-life and/or higher solubility as compared to the corresponding protein that is deficient in said URP.
4. The URP of claim 1 , wherein upon incorporation of the URP into to an N- and/or C-terminus of a heterologous protein to create a monomeric fusion protein, said heterologous protein exhibits a serum secretion half-life that is at least two times longer as compared to the corresponding protein that is deficient in said URP.
5. The URP of claim 1 , wherein one of the amino acids selected from the group consisting of glycine (G), aspartate (D), alanine (A), serine (S), threonine (T), glutamate (E) and proline (P) alone constitutes at least 20% of the total amino acids of the URP.
6. The URP of claim 1 , wherein one of the amino acids selected from the group consisting of glycine (G), aspartate (D), alanine (A), serine (S), threonine (T), glutamate (E) and proline (P) alone constitutes at least 30% of the total amino acids of the URP.
7. The URP of claim 1 , wherein the URP comprises at least 100 contiguous amino acids.
8. The URP of claim 1 , wherein the URP comprises at least 200 contiguous amino acids.
9. The URP of claim 1 , wherein the URP comprises at least 400 contiguous amino acids.
10. The URP of claim 1 , wherein any one type of amino acid is no more than 30% of the total amino acids of the URP.
11. The URP of claim 4 , wherein the heterologous protein is a growth hormone.
12. The URP of claim 11 , wherein the growth hormone is human growth hormone.
13. The URP of claim 4 , wherein the heterologous protein is a glucagon.