Method for monomerizing matrix metalloproteinase 7 (MMP-7) aggregate
A method for monomerization of MMP-7 aggregates is provided. A method for monomerization of MMP-7 aggregates which comprises treating MMP-7 aggregates with a buffer solution comprising a monovalent cation chloride (sodium chloride, potassium chloride, etc.) at a low concentration or with a buffer solution not comprising a monovalent cation chloride, a process for preparing MMP-7 which involves said method for monomerization, and a (pharmaceutical) composition comprising MMP-7 in the aforementioned buffer solution. In case that a (pharmaceutical) composition comprising MMP-7 at a low concentration is prepared, the aforementioned buffer solution comprising sugar alcohols or sugars is used.
1. A method for monomerization of matrix metalloproteinase 7 (MMP-7) aggregates which comprises treating MMP-7 aggregates with a solution comprising a monovalent cation compound at 80 mM or less or with a solution not comprising a monovalent cation compound, wherein MMP-7 monomers are obtained.
2. The method for monomerization according to claim 1 wherein the MMP-7 aggregates are treated with a solution not comprising a monovalent cation compound.
3. The method for monomerization according to claim 1 wherein the monovalent cation compound is at 40 mM or less.
4. The method for monomerization according to claim 1 wherein the monovalent cation compound is selected from the group consisting of sodium chloride, potassium chloride, sodium sulfate, potassium sulfate, sodium carbonate, potassium carbonate, sodium phosphate and potassium phosphate.
5. The method for monomerization according to claim 1 wherein the monovalent cation compound is from a monovalent cation chloride.
6. The method for monomerization according to claim 5 wherein the monovalent cation compound is selected from the group consisting of sodium chloride and potassium chloride.
7. The method for monomerization according to claim 1 wherein the solution further comprises calcium chloride.
8. The method for monomerization according to claim 7 wherein the calcium chloride is at 30 mM or less.
9. The method for monomerization according to claim 1 wherein the solution is a buffer solution.
10. The method for monomerization according to claim 9 wherein the buffer solution is 5 to 25 mM Tris buffer.
11. The method for monomerization according to claim 1 wherein the MMP-7 is at 20 mg/ml or less.
12. The method for monomerization according to claim 10 wherein the solution is 5 to 25 mM Tris buffer having a pH of 6 to 8 comprising 30 to 40 mM sodium chloride and 5 to 30 mM calcium chloride.
13. The method for monomerization according to claim 1 wherein the solution further comprises sugar alcohols and/or sugars.
14. The method for monomerization according to claim 13 wherein the sugar alcohols and/or sugars are selected from the group consisting of sucrose, lactose, maltose, xylose, trehalose, mannitol, sorbitol, xylitol, maltitol, lactitol, and oligosaccharide alcohols.
15. The method for monomerization according to claim 13 wherein the sugar alcohols and/or sugars are at 2 w/v % or more.
16. The method for monomerization according to claim 15 wherein the sugar alcohols and/or sugars are at 2 to 7 w/v %.
17. The method for monomerization according to claim 14 wherein the sugar alcohols and/or sugars are mannitol or sucrose.
18. The method for monomerization according to claim 17 wherein the mannitol is at 2 to 5% and the sucrose is at 2 to 7 w/v %.
19. A process for preparing monomers of MMP-7 comprising the following steps:
a step of disrupting cells producing proMMP-7 inclusion body;
a step of dissolution/refolding treatment of proMMP-7 inclusion body;
a step of purification of proMMP-7;
a step of self-activation of proMMP-7 into MMP-7; and
a step comprising treating MMP-7 with a solution comprising a monovalent cation compound at 80 mM or less or with a solution not comprising a monovalent cation compound, wherein MMP-7 monomers are obtained.
20. The process according to claim 19 , further comprising a step of concentration using an ultrafiltration membrane.