Compositions of GM-CSF and interleukin fusions for immune modulation and uses related thereto
This disclosure relates to recombinant proteins comprising a GM-CSF sequence and an interleukin sequence and nucleic acids related thereto. In certain embodiments, the disclosure relates to recombinant proteins comprises N-terminal sequences that are the result of improved production techniques and uses for treating or preventing autoimmune diseases such as multiple sclerosis and cancer.
1. A recombinant protein consisting of GHMAPARS 11 PS 12 PS 13 T 14 QPWEHVNAIQEARRLLN 15 LSRDTAAEMN 16 ETVEVISEMFDLQEPTCLQTRLELYKQGLRGSLTKLKGPLTMMASHYKQHCPPTPETSCATQIITFESFKENLKDFLLVGSMRISKPHLRSISIQCYLCLLLNSHFLTEAGIHVFILGCFSAGLPKTEANWVNVISDLKKIEDLIOSMHIDATLYTESDVHPSCKVTAMKCFLLELOVISLESGDASIHDTVENLIILANNSLSSNGN 17 VTESGCKECEELEEKNIKEFLQSFVHIVQMFINTS (SEQ ID NO: 12) wherein S 11 , S 12 , S 13 , T 14 , N 15 , N 16 , and N 17 are not conjugated to a glycan.
2. The recombinant protein of claim 1 consisting of SEQ ID NO: 12 made by the process of,
expressing a pre-cleavage recombinant protein in a prokaryotic cell, wherein the pre-cleavage recombinant protein comprises a cleavage sequence E-X 18 -X 19 -X 20 -Q-X 21 -H (SEQ ID NO: 13), wherein X 18 , X 19 , and X 20 are independently any amino acid, and X 21 is G, and
mixing the pre-cleavage recombinant protein with a tobacco etch virus (TEV) protease under conditions such that a recombinant protein with an N-terminal comprising X 21 -H— is formed.
3. The recombinant protein of claim 2 , wherein the cleavage sequence is ENLYFQGH (SEQ ID NO: 14).