PARTIAL PURIFICATION METHOD FOR HYDROPHILIC RECOMBINANT PROTEIN
The present invention relates to a method for partially purifying a hydrophilic recombinant protein from a host expressing a hydrophilic recombinant protein. The method includes: a solution acquisition step in which a dissolving solvent is added to the host expressing a hydrophilic recombinant protein so as to dissolve the host cell and an insoluble substance is separated therefrom so as to obtain a solution. The hydrophilic recombinant protein has a hydropathy index of 0 or less, and the dissolving solvent is an aprotic polar solvent. Preferably, the solution is subjected to solvent substitution using an aqueous solvent.
1 . A method for partially purifying a hydrophilic recombinant protein from a host expressing a hydrophilic recombinant protein,
the method comprising a solution acquisition step in which a dissolving solvent is added to the host expressing a hydrophilic recombinant protein so as to dissolve the host cell and an insoluble substance is separated therefrom so as to obtain a solution,
wherein the hydrophilic recombinant protein has a hydropathy index of 0 or less, and
the dissolving solvent is an aprotic polar solvent.
2 . The method for partially purifying a hydrophilic recombinant protein according to claim 1 , wherein the dissolving solvent has a dipole moment of 3.0 D or more.
3 . The method for partially purifying a hydrophilic recombinant protein according to claim 1 , wherein the dissolving solvent is one or more kinds of aprotic polar solvent selected from the group consisting of dimethyl sulfoxide, N,N-dimethylformamide, N,N-dimethylacetamide, 1,3-dimethyl-2-imidazolidone, and N-methyl-2-pyrrolidone.
4 . The method for partially purifying a hydrophilic recombinant protein according to claim 1 , wherein the dissolving solvent is a solvent obtained by adding an inorganic salt to the aprotic polar solvent.
5 . The method for partially purifying a hydrophilic recombinant protein according to claim 1 , wherein the separation of the insoluble substance is performed by filtration and/or centrifugation.
6 . The method for partially purifying a hydrophilic recombinant protein according to claim 1 , wherein an addition amount of the dissolving solvent is 0.5 times or more a mass of the host by volume (mL)/mass (g) ratio.
7 . The method for partially purifying a hydrophilic recombinant protein according to claim 1 , wherein the solution is subjected to solvent substitution using an aqueous solvent.
8 . The method for partially purifying a hydrophilic recombinant protein according to claim 7 , wherein the solution after solvent substitution is processed further by at least one selected from the group consisting of a column, filtration, and centrifugation.
9 . The method for partially purifying a hydrophilic recombinant protein according to claim 1 , wherein the hydrophilic recombinant protein is a protein selected from the group consisting of a recombinant spider silk protein, a recombinant resilin, and a recombinant collagen.
10 . The method for partially purifying a hydrophilic recombinant protein according to claim 2 , wherein the dissolving solvent is a solvent obtained by adding an inorganic salt to the aprotic polar solvent.
11 . The method for partially purifying a hydrophilic recombinant protein according to claim 3 , wherein the dissolving solvent is a solvent obtained by adding an inorganic salt to the aprotic polar solvent.
12 . The method for partially purifying a hydrophilic recombinant protein according to claim 2 , wherein the separation of the insoluble substance is performed by filtration and/or centrifugation.
13 . The method for partially purifying a hydrophilic recombinant protein according to claim 3 , wherein the separation of the insoluble substance is performed by filtration and/or centrifugation.
14 . The method for partially purifying a hydrophilic recombinant protein according to claim 2 , wherein an addition amount of the dissolving solvent is 0.5 times or more a mass of the host by volume (mL)/mass (g) ratio.
15 . The method for partially purifying a hydrophilic recombinant protein according to claim 2 , wherein the solution is subjected to solvent substitution using an aqueous solvent.
16 . The method for partially purifying a hydrophilic recombinant protein according to claim 3 , wherein the solution is subjected to solvent substitution using an aqueous solvent.
17 . The method for partially purifying a hydrophilic recombinant protein according to claim 4 , wherein the solution is subjected to solvent substitution using an aqueous solvent.
18 . The method for partially purifying a hydrophilic recombinant protein according to claim 10 , wherein the solution is subjected to solvent substitution using an aqueous solvent.
19 . The method for partially purifying a hydrophilic recombinant protein according to claim 15 , wherein the solution after solvent substitution is processed further by at least one selected from the group consisting of a column, filtration, and centrifugation.
20 . The method for partially purifying a hydrophilic recombinant protein according to claim 2 , wherein the hydrophilic recombinant protein is a protein selected from the group consisting of a recombinant spider silk protein, a recombinant resilin, and a recombinant collagen.