Methods of crystallizing antibodies
This invention relates to crystals of whole antibodies and fragments thereof, and formulations and compositions comprising such crystals. More particularly, methods are provided for the crystallization of high concentrations of whole antibodies, and fragments thereof, in large batches, and for the preparation of stabilized whole antibody crystals for use alone, or in dry or slurry formulations or compositions. This invention also relates to methods for stabilization, storage and delivery of biologically active whole antibody crystals. The present invention further relates to methods using whole antibody crystals, antibody fragment crystals, or compositions or formulations comprising such crystals for biomedical applications, including biological delivery to humans and animals. More particularly, whole antibody crystals or antibody fragment crystals, or crystal compositions or formulations thereof, are used as a carrier-free delivery system which can slowly release active whole antibodies or fragments thereof, to a subject, where and when they are needed.
1. In a method of crystallizing an antibody from seed crystals, the improvement comprising the steps of:
(a) providing said seed crystals of a whole antibody, wherein the antibody is selected from an immunoglobulin of class IgG, IgM, IgA, IgD, IgE, and serum IgA (sIgA);
(b) mixing at least 1 ml of a solution of the whole antibody with a crystallization solution or a crystallization buffer;
(c) adding said seed crystals of said whole antibody to the mixture obtained in step (b); and
(d) agitating said mixture for between about 3 hours and about 48 hours at room temperature, until crystals of said whole antibody are formed,
wherein said method can be used to obtain a large-batch of whole antibody crystals from an antibody concentration of between 1 mg/ml and 500 mg/ml in step (b), and at a crystallization buffer pH from 1.9 to 11.1.
2. The crystallization method according to claim 1 , further comprising the step of drying said crystals by a method selected from the group consisting of: air drying, spray drying, lyophilization, vacuum oven drying and nitrogen gas drying.
3. The crystallization method as described in claim 1 , wherein step (b) comprises mixing 1 ml of the solution of whole antibody with 1 ml of the crystallization buffer.
4. The crystallization method as described in claim 1 , wherein step (b) comprises mixing 4 ml of the solution of whole antibody with 4 ml of the crystallization buffer.
5. The crystallization method as described in claim 1 , wherein step (b) comprises mixing 8 ml of the solution of whole antibody with 8 ml of the crystallization buffer.
6. The crystallization method as described in claim 1 , wherein agitating comprises stirring.
7. The crystallization method as described in claim 1 , wherein the antibody is Rituximab.
8. The crystallization method as described in claim 1 , wherein the antibody is Trastuzumab.
9. The crystallization method as described in claim 1 , wherein the antibody is Infliximab.
10. The crystallization method as described in claim 1 , wherein the antibody is immunoglobulin class IgG.
11. The crystallization method as described in claim 10 , wherein the antibody is immunoglobulin subclass IgG1.
12. The crystallization method as described in claim 1 , wherein the antibody is selected from immunoglobulin subclass IgG1, IgG2, IgG3 and IgG4,IgM1 and IgM2, and IgA1 and IgA2.