Stable formulations of immunoglobulin single variable domains
The present invention relates to stable formulations of polypeptides, e.g. immunoglobulin single variable domains, in particular immunoglobulin single variable domains directed against von Willebrand Factor (vWF). The invention provides formulations which are stable upon storage for prolonged periods of time and over a broad range of temperatures. The formulations of the invention ensure a high stability of the polypeptide, allowing multiple freeze-thaw cycles without chemical or physical deterioration, and provide stability in relation to mechanical stress, such as shake, shear or stir stress. They are suitable for pharmaceutical and diagnostic preparations and compatible with pharmaceutically acceptable diluents.
1 . A formulation comprising a von Willebrand Factor (vWF) binder, a citrate buffer, an excipient, and polysorbate 80, wherein:
(a) the vWF binder has a concentration from about 0.1 mg/mL to about 80 mg/mL;
(b) the excipient is sucrose at a concentration of about 5% to about 15% (w/v), wherein the sucrose is not at a concentration of about 7% (w/v);
(c) the polysorbate 80 has a concentration of about 0.001% to 0.5% (v/v); and
(d) the citrate buffer has a concentration of about 5 mM to about 200 mM such that the pH of the formulation is about 6.0 to about 7.0,
wherein said vWF binder comprises at least one of SEQ ID NOs: 1-19.
2 . The formulation according to claim 1 , which has:
(i) less than 5% of high molecular weight (HMW) species after storage for at least 12 months at 5° C.; and/or
(ii) less than 5% of low molecular weight (LMW) species after storage for at least 12 months at 5° C.
3 . The formulation according to claim 1 , which is a liquid or reconstituted lyophilized formulation.
4 . The formulation according to claim 1 , which is in a lyophilized, spray-dried, or frozen form.
5 . The formulation according to claim 1 , wherein said vWF binder comprises an amino acid sequence which is at least 90% identical to SEQ ID NO: 1.
6 . The formulation according to claim 1 , wherein the vWF binder in the formulation retains at least 80% of its stability after storage for at least 12 months at 5° C.
7 . The formulation according to claim 1 , wherein at least 90% of the vWF binder retains its binding activity after storage compared to the binding activity prior to storage, said binding activity as measured by ELISA and/or surface plasmon resonance.
8 . The formulation according to claim 1 , wherein at least 95% of the vWF binder retains its binding activity after storage compared to the binding activity prior to storage, said binding activity as measured by ELISA and/or surface plasmon resonance.
9 . The formulation according to claim 1 , wherein at least 99% of the vWF binder retains its binding activity after storage compared to the binding activity prior to storage, said binding activity as measured by ELISA and/or surface plasmon resonance.
10 . The formulation according to claim 1 , wherein said vWF binder comprises an amino acid sequence which is identical to SEQ ID NO: 1.
11 . An article of manufacture comprising a container comprising the formulation according to claim 1 , which is a bulk storage formulation, wherein the container is suitable for storing at least 100 liters of the formulation at below freezing conditions.
12 . A kit or an article of manufacture, comprising a container containing the formulation of claim 1 , and instructions for use.
13 . The kit or article of manufacture of claim 12 , wherein the formulation is present in a vial or an injectable syringe.
14 . A formulation comprising a von Willebrand Factor (vWF) binder, a citrate buffer, an excipient, and polysorbate 80, wherein:
(a) the vWF binder has a concentration from about 0.1 mg/mL to about 80 mg/mL;
(b) the excipient is sucrose at a concentration of about 5% to about 15% (w/v);
(c) the polysorbate 80 has a concentration of about 0.001% to 0.5% (v/v), wherein the polysorbate 80 does not have a concentration of about 0.01% (w/v); and
(d) the citrate buffer has a concentration of about 5 mM to about 200 mM such that the pH of the formulation is about 6.0 to about 7.0,
wherein said vWF binder comprises at least one of SEQ ID NOs: 1-19.
15 . The formulation of claim 14 , wherein said vWF binder comprises the amino acid sequence of SEQ ID NO: 1.
16 . The formulation according to claim 14 , which is a liquid or reconstituted lyophilized formulation.
17 . A formulation comprising a von Willebrand Factor (vWF) binder, a citrate buffer, an excipient, and polysorbate 80, wherein:
(a) the vWF binder has a concentration from about 0.1 mg/mL to about 80 mg/mL;
(b) the excipient is sucrose at a concentration of about 5% to about 15% (w/v);
(c) the polysorbate 80 has a concentration of about 0.001% to 0.5% (v/v); and
(d) the citrate buffer has a concentration of about 5 mM to about 200 mM such that the pH of the formulation is about 6.0 to about 7.0, wherein the citrate buffer does not have a concentration of about 20 mM,
wherein said vWF binder comprises at least one of SEQ ID NOs: 1-19.
18 . The formulation of claim 17 , wherein said vWF binder comprises the amino acid sequence of SEQ ID NO: 1.
19 . The formulation according to claim 17 , which is a liquid or reconstituted lyophilized formulation.