Composition comprising recombinant clostridium neurotoxin
The invention provides methods for producing soluble di-chain BoNT/A protein.
1. A liquid pharmaceutical composition comprising:
an active di-chain BoNT/A protein;
Lys-C;
a non-protein stabilizing agent that is a surfactant; and
water;
wherein:
the composition does not comprise a protein stabilizing agent;
the Lys-C is present at a concentration of less than 400 pg Lys-C per 100 ng BoNT/A protein; and
less than 2% of the BoNT/A in the composition is single-chain BoNT/A.
2. The liquid pharmaceutical composition of claim 1 , wherein the composition contains Lys-C at a concentration of less than 300 pg Lys-C per 100 ng BoNT/A protein.
3. The liquid pharmaceutical composition of claim 1 , further comprising:
sodium chloride;
a buffer to maintain pH between 5.5 and 7.5; and
a disaccharide;
wherein the water is sterile water.
4. The liquid pharmaceutical composition of claim 2 , wherein the composition contains Lys-C at a concentration of less than 200 pg Lys-C per 100 ng BoNT/A protein.
5. The liquid pharmaceutical composition of claim 2 , wherein the composition contains Lys-C at a concentration of less than 100 pg Lys-C per 100 ng BoNT/A protein.
6. The liquid pharmaceutical composition of claim 2 , wherein the composition contains Lys-C at a concentration of less than 50 pg Lys-C per 100 ng BoNT/A protein.
7. The liquid pharmaceutical composition of claim 2 , wherein the composition contains Lys-C at a concentration of less than 20 pg Lys-C per 100 ng BoNT/A protein.
8. The liquid pharmaceutical composition of claim 2 , wherein the composition contains Lys-C at a concentration of less than 10 pg Lys-C per 100 ng BoNT/A protein.
9. The liquid pharmaceutical composition of claim 1 , wherein less than 1% of the BoNT/A in the composition is single-chain BoNT/A.
10. The liquid pharmaceutical composition of claim 1 , produced using a method wherein a soluble single-chain BoNT/A protein is contacted with Lys-C in solution and, following such contact, the BoNT/A is separated from Lys-C by contacting the solution containing the BoNT/A and Lys-C with a hydrophobic surface, wherein the BoNT/A binds with preference to the hydrophobic surface.
11. The liquid pharmaceutical composition of claim 3 consisting of:
BoNT/A protein wherein less than 2% of the protein is in single-chain form;
a non-protein stabilizing agent that is a surfactant;
water;
Lys-C at less than 400 pg Lys-C per 100 ng BoNT/A protein;
sodium chloride;
a buffer to maintain pH between 5.5 and 7.5; and
a disaccharide.