Recombinant proteins based on fibrinogen
The present invention relates to isolated, recombinant polypeptides comprising the gamma chain of fibrinogen and a His-Tag. The present invention further relates to nucleic acids encoding said polypeptides, vectors comprising said nucleic acids, host cells, preferably bacterial host cells, comprising said nucleic acids or vectors, biohybrid materials comprising conjugates comprising said polypeptides, uses of the above, and methods for producing said polypeptides.
1 . An isolated, recombinant polypeptide, comprising
(i) the gamma chain of fibrinogen, and
(ii) a His-Tag;
wherein said polypeptide comprises
(a) the amino acid sequence according to SEQ ID NO:3, or
(b) an amino acid sequence having at least 95% sequence identity to SEQ ID NO: 3 and having fibrinogen gamma chain activity and His-Tag functionality;
wherein fibrinogen gamma chain activity refers to the characteristic of having functional binding sites for integrins, leukocytes, platelets, fibroblasts, endothelial cells and/or fibroblast growth factor 2 (FGF-2).
2 . The isolated, recombinant polypeptide according to claim 1 , wherein said gamma chain of fibrinogen comprises binding sites for integrins, leukocytes, platelets, fibroblasts, endothelial cells and/or fibroblast growth factor 2 (FGF-2).
3 . The isolated, recombinant polypeptide according to claim 1 , wherein said gamma chain of fibrinogen comprises
(i) the amino acid sequence according to SEQ ID NO: 1, or
(ii) an amino acid sequence having at least 95% sequence identity to SEQ ID NO: 1 and having fibrinogen gamma chain activity.
4 . The isolated, recombinant polypeptide according to claim 1 , wherein said His-Tag comprises
(i) the amino acid sequence according to SEQ ID NO: 2, or
(ii) an amino acid sequence having at least 95% sequence identity to SEQ ID NO: 2 and having His-Tag functionality.
5 . A biohybrid material, comprising a conjugate comprising the polypeptide according to claim 1 and one or more moieties, selected from the group consisting of polyethylene glycol (PEG), PEG derivatives, PEG diacrylate (PEG-DA), poly(oligo (ethylene glycol) methyl ether methacrylate (POEGMA), poly(N-(2-hydroxypropyl)-methacrylamide) (PHPMA) and HPMA copolymers, poly(vinylpyrrolidone) (PVP), poly(ethyleneimine) (PEI), poly(acroloylmorpholine) (PAcM), poly(2-ethyl 2-oxazoline) (PEOZ), divinylethermaleic anhydride/acid copolymer (DIVEMA), poly(styrene-co-maleic acid/anhydride) (SMA), poly(vinyl alcohol) (PVA) and temperature-sensitive polymers including poloxamers, poly(N-isopropylacrylamide) (PNIPAM), poly(N,N-diethylacrylamide) (PDEAM), poly(methylvinylether) (PMVE), poly(N-vinylcaprolactam) (PNVC1), and mixtures thereof.
6 . The isolated, recombinant polypeptide according to claim 1 , wherein said polypeptide comprises the amino acid sequence according to SEQ ID NO:3.
7 . The isolated, recombinant polypeptide according to claim 1 , wherein said polypeptide comprises an amino acid sequence having at least 95% sequence identity to SEQ ID NO:3 and having fibrinogen gamma chain activity and His-Tag functionality.
8 . The isolated, recombinant polypeptide according to claim 3 , wherein said gamma chain of fibrinogen comprises the amino acid sequence according to SEQ ID NO: 1.
9 . The isolated, recombinant polypeptide according to claim 3 , wherein said gamma chain of fibrinogen comprises an amino acid sequence having at least 95% sequence identity to SEQ ID NO: 1 and having fibrinogen gamma chain activity.
10 . The isolated, recombinant polypeptide according to claim 4 , wherein said His-Tag comprises the amino acid sequence according to SEQ ID NO: 2.
11 . The isolated, recombinant polypeptide according to claim 4 , wherein said His-Tag comprises an amino acid sequence having at least 95% sequence identity to SEQ ID NO: 2 and having His-Tag functionality.
12 . The isolated, recombinant polypeptide according to claim 7 , wherein said polypeptide comprises an amino acid sequence having at least 96% sequence identity to SEQ ID NO:3 and having fibrinogen gamma chain activity and His-Tag functionality.
13 . The isolated, recombinant polypeptide according to claim 12 , wherein said polypeptide comprises an amino acid sequence having at least 97% sequence identity to SEQ ID NO:3 and having fibrinogen gamma chain activity and His-Tag functionality.
14 . The isolated, recombinant polypeptide according to claim 13 , wherein said polypeptide comprises an amino acid sequence having at least 98% sequence identity to SEQ ID NO:3 and having fibrinogen gamma chain activity and His-Tag functionality.
15 . The isolated, recombinant polypeptide according to claim 14 , wherein said polypeptide comprises an amino acid sequence having at least 99% sequence identity to SEQ ID NO:3 and having fibrinogen gamma chain activity and His-Tag functionality.