Process for purifying recombinant
View Patent ↗The present invention relates to processes for purifying high-quality recombinant Plasmodium falciparum circumsporozoite protein at high yields.
1. A stable liquid formulation of recombinant P. falciparum circumsporozoite protein (rCSP), the rCSP comprising the most N-terminal cysteine, wherein the most N-terminal cysteine corresponds to the cysteine at position 25 as set forth in SEQ ID NO: 1, the formulation comprising about 1 to about 50 mg/ml rCSP in a formulation buffer comprising about 0.5 to about 1.5 mM monothioglycerol (MTG) and about 10% to about 20% arginine.
2. The stable liquid formulation of claim 1 , wherein the formulation buffer comprises 0.5× or 1×PBS.
3. The stable liquid formulation of claim 1 , wherein the formulation buffer has a pH of about 6.0 to about 7.5.
4. The stable liquid formulation of claim 2 , wherein the formulation buffer has a pH of about 6.0 to about 7.5.
5. The stable liquid formulation of claim 1 , wherein the storage temperature is about 4° C. to about 15° C.
6. The stable liquid formulation of claim 2 , wherein the storage temperature is about 4° C. to about 15° C.
7. The stable liquid formulation of claim 3 , wherein the storage temperature is about 4° C. to about 15° C.
8. The stable liquid formulation of claim 4 , wherein the storage temperature is about 4° C. to about 15° C.
9. The stable liquid formulation of claim 7 , wherein the stable liquid formulation comprises about 1 to about 5 mg/ml rCSP, about 1.0 mM MTG, about 10% arginine, 1×PBS, has a pH of about 6.0 to about 7.5, and wherein the storage temperature is about 4° C. to about 6° C.
10. The stable liquid formulation of claim 1 , wherein the stable liquid P. falciparum circumsporozoite protein formulation contains at least one of the following: not more than about 10% P. falciparum circumsporozoite protein dimer; not more than about 10% high P. falciparum circumsporozoite protein molecular weight aggregates, and; not more than about 10% P. falciparum circumsporozoite protein degradation products.
11. A stable liquid formulation of rCSP, the rCSP comprising the most N-terminal cysteine, wherein the most N-terminal cysteine corresponds to the cysteine at position 25 as set forth in SEQ ID NO: 1, the formulation comprising about 1 to about 50 mg/ml rCSP, in a formulation buffer comprising about 0.5 to about 1.5 mM monothioglycerol (MTG) and about 1% to about 20% arginine.
12. A method for stably maintaining rCSP in a stable liquid formulation, the method comprising providing the stable liquid formulation of claim 1 , in a formulation buffer of 1×PBS at a pH of about 6.4 to about 7.2, wherein the rCSP is stably maintained at a temperature of about 3° C. to about 25° C., for at least about 7 days.
13. The method of claim 12 , wherein the formulation buffer is 1×PBS at a pH of about 6.4 to about 7.0.
14. A method for stably maintaining rCSP in a stable liquid formulation, the method comprising providing the stable liquid formulation of claim 11 , in a formulation buffer of 1×PBS at a pH of about 6.4 to about 7.2, wherein the rCSP is stably maintained at a temperature of about 3° C. to about 25° C., for at least about 7 days.