1 . A method for preparing a lyophilized cake, comprising: subjecting a formulation including at least one solvent molecule and at least one peptide or protein to lyophilization to obtain a lyophilized cake, wherein said lyophilization includes a secondary drying step conducted at a temperature below about 0° C.
2 . The method of claim 1 , wherein said at least one peptide or protein comprises an antibody, an antibody fragment, a Fab region of an antibody, an antibody-drug conjugate, a fusion protein, a protein pharmaceutical product, or a drug.
3 . The method of claim 1 , wherein said lyophilized cake includes said at least one solvent molecule at a target weight percentage.
4 . The method of claim 1 , wherein a weight percentage of said at least one solvent molecule in said lyophilized cake is controlled by said secondary drying step.
5 . The method of claim 1 , wherein a weight percentage of said at least one solvent molecule in said lyophilized cake is about 3-5%, about 4%, or about 4.5%.
6 . The method of claim 1 , wherein said lyophilized cake is stable in room temperature storage and/or has improved stability for refrigerated storage.
7 . The method of claim 1 , wherein said at least one solvent molecule comprises a water molecule.
8 . The method of claim 1 , wherein said lyophilization further comprises determining an ending of a primary drying step based on a change of a pressure in the chamber of the freeze-dryer.
9 . The method of claim 1 , wherein a temperature of said secondary drying step is about −10° C., about −20° C., or about −30° C.
10 . The method of claim 1 , wherein a temperature of said secondary drying step is about the same as or lower than a temperature of a first drying step.
11 . The method of claim 1 , wherein a duration of said secondary drying step is about 10 to about 150 hours, about 10 hours, about 30 hours, about 50 hours, about 100 hours, or about 150 hours.
12 . The method of claim 1 , wherein said formulation further comprises at least one buffer, at least one excipient, at least one stabilizer, at least one cryo-protectant, at least one bulking agent, at least one plasticizer, or a combination thereof.
13 . The method of claim 12 , wherein said at least one buffer comprises acetate and/or histidine hydrochloride.
14 . The method of claim 12 , wherein said at least one buffer has a pH value of about 5.3 to about 6.
15 . The method of claim 12 , wherein said at least one excipient comprises polysorbate 80.
16 . The method of claim 12 , wherein said at least one stabilizer comprises sucrose.
17 . The method of claim 16 , wherein a ratio of said sucrose to said at least one peptide or protein is about 1:1 to 10:1, about 1:1, about 3:1, or about 10:1.
18 . The method of claim 12 , wherein said at least one stabilizer comprises a polyol, sucrose, mannitol, trehalose, sorbitol, an amino acid, or a combination thereof.
19 . The method of claim 12 , wherein said at least one cryo-protectant comprises a surfactant, sugar, salt, an amino acid, or a combination thereof.
20 . The method of claim 12 , wherein said formulation comprises: a. sucrose at a concentration of about 0% to about 5%, about 2.5%, or about 5%; b. arginine at a concentration of about 0% to about 2%, about 1%, or about 2%; c. acetate at a concentration of about 0 mM to about 10 mM, or about 10 mM; d. histidine; and/or e. polysorbate 80 at a concentration of about 0% to about 0.2%, or about 0.2%.
