Annelid haemoglobin lyophilisation process

Abstract

The present invention relates to a lyophilizate comprising at least one globin, one globin protomer or one extracellular hemoglobin of annelids, and a stabilizer chosen from disaccharides, polyols and antioxidants. The present invention also relates to a composition comprising: a solution comprising at least one globin, one globin protomer or one extracellular hemoglobin of annelids, and a stabilizer chosen from disaccharides, polyols and antioxidants. Finally, the present invention relates to a process for preparing the lyophilizate.

1 . A lyophilizate comprising at least one extracellular hemoglobin, globin protomer or globin of annelids, and a stabilizer chosen from disaccharides, polyols and antioxidants. 
     
     
         2 . A composition comprising:
 a solution comprising at least one extracellular hemoglobin, globin protomer or globin of annelids, and   a stabilizing chosen from trehalose and ascorbic acid.   
     
     
         3 . The lyophilizate of  claim 1 , wherein the extracellular hemoglobin of annelids is an extracellular hemoglobin of polychaete annelids. 
     
     
         4 . The lyophilizate of  claim 1 , wherein the stabilizer is chosen from trehalose, sucrose, mannitol, sorbitol and ascorbic acid, preferably from trehalose and ascorbic acid. 
     
     
         5 . A process for preparing the lyophilizate of  claim 1 , comprising:
 i) the mixing of a solution comprising at least one extracellular hemoglobin, globin protomer or globin of annelids with a stabilizer chosen from disaccharides, polyols and antioxidants;   ii) the freezing of the mixture at a temperature of between −10° C. and −100° C. for a time of at least 24 h;   iii) the sublimation of the frozen mixture for at least 2 h, under vacuum;   iv) the final drying of the mixture until a powder is obtained.   
     
     
         6 . The process of  claim 5 , wherein the stabilizer comprises disaccharides or polyols in a concentration of between 1 and 500 mg/ml. 
     
     
         7 . The process of  claim 5 , wherein the stabilizer comprises antioxidants in a concentration of between 3 and 20 mM. 
     
     
         8 . The process of  claim 5 , wherein the stabilizer comprises trehalose or sucrose present in a concentration of between 50 and 70 mg/ml, mannitol present in a concentration of approximately 50 to 100 g/l and ascorbic acid present in a concentration of approximately 5 mM. 
     
     
         9 . The process of  claim 5 , wherein the freezing is carried out at a temperature of between −10° C. and −90° C., for at least 24 h. 
     
     
         10 . The process of  claim 5 , wherein the sublimation is carried out for at least 4 h.