Spacers to increase the expression of recombinant fusion proteins
The present invention relates to fusion proteins. The invention specifically relates to compositions and methods of Tf-based fusion proteins that demonstrate a high-level expression of transferrin (Tf)-based fusion proteins by inserting a helical linker between two protein domains.
1. A two-domain fusion protein construct for improving expression of a fusion protein having a first protein domain and a second protein domain, said construct comprising:
an alpha-helical peptide linker inserted between said first and second domains of the fusion protein,
wherein inclusion of said alpha-helical peptide linker in said fusion protein increases the expression level of said fusion protein as compared to fusion proteins comprising said two domains without said linker.
2. The fusion protein construct according to claim 1 , wherein said alph-helical peptide linker comprises more than one helical motif between said first and second domains.
3. The fusion protein construct according to claim 1 , wherein said alpha-helical peptide linker comprises LEA(EAAAK) 4 ALE (SEQ ID NO: 1).
4. The fusion protein construct according to claim 1 , wherein:
said first protein domain comprises a carrier protein selected from the group consisting of a transferrin, a serum albumin, an antibody and sFv; and
said second protein domain comprises a therapeutic protein selected from the group consisting of a colony stimulating factor (CSF), an interferon, a cytokine, a hormone, a lymphokine, an interleukin, a hematopoietic growth factor, and a toxin.
5. The fusion protein construct according to claim 4 , wherein said second protein domain comprises a granulocyte colony stimulating factor (G-CSF) or a human growth hormone (hGH).
6. The fusion protein construct according to claim 1 , wherein said first protein domain is a transferrin (Tf)-based protein domain.