Methods of preventing aggregation of various substances upon rehydration or thawing and compositions obtained thereby
The present invention encompasses methods to prevent aggregation of a wide variety of substances during freezing/thawing and/or dehydrating/rehydrating. The substances thus obtained and compositions comprising the substances are also encompassed by the invention.
1 . A method of reducing aggregation during dehydration and rehydration of substances comprising the steps of
adding to a solution or suspension of the substances an amount of trehalose sufficient to prevent aggregation upon rehydration; and
dehydrating the solution or suspension.
2 . The method according to claim 1 wherein the substances are selected from the group consisting of therapeutic, prophylactic and diagnostic.
3 . The method according to claim 2 wherein the substances are therapeutic and are biological modifiers.
4 . The method according to claim 3 wherein the biological modifier is selected from the group consisting of proteins and peptides, steroid hormones, oligosaccharides, nucleic acids and small molecules.
5 . The method according to claim 4 wherein the proteins are selected from the group consisting of growth hormones, growth factors, insulin, monoclonal antibodies, interleukins and interferons.
6 . The method according to claim 5 wherein the substance is human growth hormone.
7 . The method according to claim 4 wherein the steroid hormones are selected from the group consisting of estrogen, progesterone and testosterone.
8 . The method according to claim 2 wherein the substances are prophylactic substances and are aluminum based adjuvants.
9 . The method according to claim 8 further comprising the step of incorporating the adjuvants into vaccines.
10 . The method according to claim 9 wherein the vaccines are diphtheria/tetanus/pertussis (DTP) or inactivated poliovaccine.
11 . The method according to claim 10 wherein the vaccine is DTP.
12 . The method according to claim 2 wherein the substance is diagnostic and is selected from the group consisting of colloidal gold, polystyrene latex, fixed erythrocytes and monoclonal antibodies.
13 . The method according to claim 12 wherein the substance is red blood cells, further comprising the step of fixing the red blood cells prior to adding trehalose.
14 . The method according to claim 13 wherein the fixing is by glutaraldehyde.
15 . The method according to claim 1 wherein the trehalose is added in an amount to obtain a final concentration of from about 1% to 50% (w/v).
16 . The method according to claim 1 wherein the trehalose is added in an amount to obtain a final concentration of from about 5% to 25% (w/v).
17 . The method according to claim 1 wherein the dehydration step occurs by lyophilization, drying at ambient conditions or drying under reduced vapor pressure.
18 . The method according to claim 17 wherein the drying at reduced vapor pressure occurs at room temperature or at a temperature elevated above room temperature but below a temperature at which degradation or chemical change of the substance occurs.
19 . The method according to claim 1 further comprising the step of rehydrating the substance to obtain a solution or suspension of substantially nonaggregated substance.
20 . A method of reducing aggregation of substances in solution or suspension during freezing comprising the steps of:
adding to the solution or suspension of the substance an amount of trehalose sufficient to prevent aggregation during freezing; and
freezing the solution or suspension.
21 . The method according to claim 18 wherein the substances are selected from the group consisting of therapeutic, prophylactic and diagnostic.
22 . The method according to claim 21 wherein the substance is therapeutic and is a biological response modifier.
23 . The method according to claim 22 wherein the biological modifier is selected from the group consisting of proteins and peptides, steroid hormones, oligosaccharides, nucleic acids and small molecules.
24 . The method according to claim 23 wherein the proteins are selected from the group consisting of growth hormones, growth factors, insulin, monoclonal antibodies, interleukins and interferons.
25 . The method according to claim 24 wherein the substance is human growth hormone.
26 . The method according to claim 23 wherein the steroid hormones are selected from the group consisting of estrogen, progesterone and testosterone.
27 . The method according to claim 21 wherein the substances are prophylactic substances and are aluminum based adjuvants.
28 . The method according to claim 27 further comprising the step of incorporating the adjuvants into vaccines.
29 . The method according to claim 28 wherein the vaccines are diphtheria/tetanus/pertussis (DTP) or diphtheria/tetanus/pertussis/inactivated poliovaccine (DTP/IPV).
30 . The method according to claim 29 wherein the vaccine is DTP/IPV.
31 . The method according to claim 20 wherein the substance is diagnostic and is selected from the group consisting of colloidal gold, polystyrene latex, fixed erythrocytes and monoclonal antibodies.
32 . The method according to claim 20 wherein the trehalose is added in an amount to attain of from about 1% to 50% (w/v).
33 . The method according to claim 32 wherein the trehalose is added in an amount to attain of from about 5% to 25% (w/v).
34 . The method according to claim 20 further comprising the step of thawing the solution or suspension to obtain a solution or suspension of substantially nonaggregated substance.
35 . An aqueous composition comprising a substance and an amount of trehalose sufficient to prevent substantial aggregation of the substance upon freezing and thawing or dehydrating and rehydrating.
36 . A frozen composition comprising a substance and an amount of trehalose sufficient to prevent substantial aggregation of the substance upon thawing.
37 . A dehydrated composition comprising a substance and an amount of trehalose sufficient to prevent aggregation of the substance upon rehydration.
38 . A composition obtained by the method according to claim 1 .
39 . A composition obtained by the method according to claim 13 .
40 . A composition obtained by the method according to claim 19 .
41 . A composition obtained by the method according to claim 20 .
42 . A composition obtained by the method according to claim 34.