Means and methods of sterilization of biofunctional compositions

Abstract

The present invention, inter alia, relates to a closed sterilized container comprising at least one carrier which is a stabilizer; and at least one biomolecule reversibly attached to the carrier, wherein said carrier partially or completely covers the attached biomolecules and wherein said at least one carrier is selected from the group consisting of (poly)peptides such as dipeptides or tripeptides, amino acids, polyalcohols, polyethyleneglycols, ionic liquids, compatible solutes, saponins and a mixture thereof. The invention also relates to methods for producing sterilized containers according to the invention and uses thereof.

1 - 26 . (canceled) 
     
     
         27 . A sterile container selected from vials, ampullas, cryocontainers, phials, flasks, bottles and bags, comprising
 a carrier, and   at least one biomolecule, wherein the at least one biomolecule is a biomolecule of an eukaryotic cell, a prokaryotic cell, a virus, or a capsid, or is a fragment of said biomolecule and wherein the at least one biomolecule is reversibly attached to the carrier,   wherein the carrier   comprises a stabilizer composition comprising at least three different naturally occurring amino acids, wherein the at least three different naturally occurring amino acids are not in the form of peptides, and   partially or completely covers the attached biomolecules.   
     
     
         28 . The container according to  claim 27 , wherein the at least one biomolecule is attached to the carrier such that it can be released from the carrier prior to use. 
     
     
         29 . The container according to  claim 27 , wherein the carrier is solid. 
     
     
         30 . The container according to  claim 27 , wherein the carrier is semi-solid. 
     
     
         31 . The container according to  claim 27 , wherein the carrier is solubilizable. 
     
     
         32 . The container according to  claim 29 , wherein the solid carrier is porous. 
     
     
         33 . The container according to  claim 31 , wherein the solubilizable carrier comprises proteinaceous and/or carbohydrate structures that dissolve in aqueous, optionally buffered solutions. 
     
     
         34 . The container according to  claim 27 , wherein the at least one stabilizer is comprised in a buffered solution. 
     
     
         35 . The container according to  claim 27 , wherein the stabilizer composition comprises a mixture of at least four different naturally occurring amino acids. 
     
     
         36 . The container according to  claim 35 , wherein the mixture comprises at least one amino acid of each group of
 (a) an amino acid with nonpolar, aliphatic R groups;   (b) an amino acid with polar, uncharged R groups;   (c) an amino acid with positively charged R groups;   (d) an amino acid with negatively charged R groups; and   (e) an amino acid with aromatic R groups.   
     
     
         37 . The container according to  claim 35 , wherein the amino acids comprised in said mixture are alanine, glutamic acid, lysine, threonine, and tryptophan. 
     
     
         38 . The container according to  claim 35 , wherein the amino acids comprised in said mixture are aspartate, arginine, phenylalanine, serine, and valine. 
     
     
         39 . The container according to  claim 27 , wherein the container is a closable sterile container. 
     
     
         40 . A method for producing the sterile container according to  claim 27 , comprising:
 (a) reversibly attaching at least one biomolecule to a carrier,
 (i) wherein the carrier: 
 comprises a stabilizer composition comprising at least three different naturally occurring amino acids, wherein the at least three different naturally occurring amino acids are not in the form of peptides, and 
 partially or completely covers the attached biomolecules, and 
 (ii) wherein the at least one biomolecule comprises: 
 a biomolecule of an eukaryotic cell, a prokaryotic cell, a virus, or a capsid, or is a fragment of said biomolecule, or 
   an antibody, enzyme, receptor, membrane protein, transport protein, blood coagulation factor, hormone, cytokine and function fragments of any of the foregoing;   (b) inserting the at least one biomolecule reversibly attached to the carrier into a container;   (c) incubating the at least one biomolecule in a solution comprising at least one stabilizer selected from (poly)peptides, amino acids, carbohydrates, polyalcohols, polyethyleneglycols, ionic liquids, compatible solutes, saponins, or a mixture thereof;   (d) sealing the container with a lid; and   (e) sterilizing the container.   
     
     
         41 . The method according to  claim 40  wherein reversibly attaching of the biomolecule to the carrier and/or incubating in the solution comprising at least one stabilizer is performed in bulk production, optionally comprising a cutting step prior to the insertion of the coated carrier into the container. 
     
     
         42 . The method according to  claim 40 , wherein reversibly attaching comprises drying the carrier together with the biomolecule. 
     
     
         43 . The method according to  claim 40 , wherein sterilizing is performed by ethyleneoxide (EO), beta radiation, gamma radiation, X-ray, heat inactivation, autoclaving, or plasma sterilization. 
     
     
         44 . The method according to  claim 40 , wherein the stabilizer composition comprises a mixture of at least four different naturally occurring amino acids. 
     
     
         45 . The method according to  claim 44 , wherein the mixture comprises at least one amino acid of each group of
 (a) an amino acid with nonpolar, aliphatic R groups;   (b) an amino acid with polar, uncharged R groups;   (c) an amino acid with positively charged R groups;   (d) an amino acid with negatively charged R groups; and   (e) an amino acid with aromatic R groups.   
     
     
         46 . The method according to  claim 44 ,
 wherein the amino acids comprised in said mixture are alanine, glutamic acid, lysine, threonine, and tryptophan; or   wherein the amino acids comprised in said mixture are aspartate, arginine, phenylalanine, serine, and valine.