Interferon-alpha polypeptides and conjugates
View Patent ↗The present invention provides interferon-alpha polypeptides and conjugates, and nucleic acids encoding the polypeptides. The invention also includes compositions comprising these polypeptides, conjugates, and nucleic acids; cells containing or expressing the polypeptides, conjugates, and nucleic acids; methods of making the polypeptides, conjugates, and nucleic acids; and methods of using the polypeptides, conjugates, and nucleic acids.
1. An isolated or recombinant polynucleotide comprising a nucleic acid sequence which encodes a polypeptide having the sequence SEQ ID NO:13 or a sequence which differs from SEQ ID NO:13 in 1-8 amino acid positions, which comprises Ala or Leu at position 48 and optionally comprises a methionine at the N-terminus, which polypeptide exhibits antiviral activity.
2. The polynucleotide of claim 1 , comprising a nucleic acid sequence which encodes a polypeptide comprising the sequence SEQ ID NO:13.
3. The polynucleotide of claim 2 , comprising a nucleic acid sequence selected from SEQ ID NO:62, SEQ ID NO:63, SEQ ID NO:64, and SEQ ID NO:89.
4. An isolated host cell comprising the nucleic acid of claim 1 .
5. A vector comprising the polynucleotide of claim 1 .
6. The vector of claim 5 , which is an expression vector comprising the polynucleotide operably linked to a promoter.
7. An isolated host cell comprising the vector of claim 6 .
8. A composition comprising the polynucleotide of claim 1 and an excipient.
9. A method for preparing a polypeptide, the method comprising:
providing a culture comprising a host cell, the host cell comprising an expression vector comprising a promoter operably linked to the polynucleotide of claim 1 ,
culturing the culture under conditions which permit expression of the polypeptide, and
recovering the polypeptide.
10. The method of claim 9 , wherein the host cell is a glycosylating host cell.
11. The method of claim 9 , wherein the host cell is a bacterial host cell.
12. The method of claim 9 , wherein the bacterial host cell is E. coli.
13. The method of claim 12 , wherein the step of recovering the polypeptide comprises:
(a) isolating inclusion bodies (IB) comprising the polypeptide;
(b) solubilizing the IB, thereby obtaining unfolded polypeptide;
(c) refolding the unfolded polypeptide, thereby obtaining refolded polypeptide; and
(d) purifying the refolded polypeptide.