COMPOSITIONS AND METHODS FOR LYOPHILIZATION OF BACTERIA OR LISTERIA STRAINS
Methods and compositions are provided for lyophilization of bacteria or Listeria strains, such as Listeria monocytogenes . Provided are methods for producing a lyophilized composition comprising a bacteria or Listeria strain, formulations for lyophilization comprising a bacteria or Listeria strain, lyophilized bacteria or Listeria strains, and methods of preparing frozen bacteria or Listeria strains for lyophilization.
1 . A method of preparing a frozen Listeria strain for lyophilization, comprising thawing the frozen Listeria strain at a temperature between about 20° C. and about 37° C.
2 . The method of claim 1 , wherein the temperature is between about 32° C. and about 37° C.
3 . The method of claim 2 , wherein the temperature is about 37° C.
4 . The method of claim 1 , wherein the frozen Listeria strain is thawed for no more than 8 hours.
5 . The method of claim 1 , wherein the frozen Listeria strain is held at about 2° C. to about 8° C. for no more than 24 hours after thawing.
6 . The method of claim 1 , wherein the frozen Listeria strain is thawed in a formulation comprising a buffer and sucrose.
7 . The method of claim 6 , wherein the formulation comprises about 1% to about 5% w/v sucrose.
8 . The method of claim 7 , wherein the formulation comprises about 2% to about 3% w/v sucrose.
9 . The method of claim 8 , wherein the formulation comprises about 2.5% w/v sucrose.
10 . The method of claim 6 , wherein the formulation does not comprise one or more of trehalose, mono sodium glutamate (MSG), and recombinant human serum albumin (rHSA).
11 . The method of claim 10 , wherein the formulation does not comprise trehalose, MSG, or rHSA.
12 . The method of claim 6 , wherein the Listeria strain is a recombinant Listeria monocytogenes strain.
13 . The method of claim 6 , wherein the Listeria strain is a recombinant Listeria monocytogenes strain,
wherein the formulation comprises about 2% to about 3% w/v sucrose, and
wherein the formulation does not comprise trehalose, MSG, or rHSA.