Methods of inactivation of viruses using n-methylglucamide and its derivatives

Abstract

This disclosure relates to methods for use in inactivating viruses. The methods of inactivating viruses with N-methylglucamides is applicable to the purification process of biologically-active drugs such as protein subunits, proteins (enzymes, factors, etc.), recombinant proteins, antibodies, vaccine or gene therapeutic products. The detergents used in this method are based on multiple N-methylglucamide homologs, consisting of a hydrophilic glucose moiety and hydrophobic fatty acid tail, linked by an amide bond. Additionally, these sugar-based detergents are nonionic by nature, which do not disrupt the drug protein, plasma biologics, non-enveloped viral vaccine or adeno associated viral particles.A method of purifying a biological product solution of interest having an unidentified enveloped virus contaminant, including incubating a biological product solution of interest with a standard solution, inactivating any potential enveloped virus contaminant present in the biological product solution of step (a), measuring the inactivated virus present in the final solution of step (b), incubating a separate biological product solution of interest with a N-methylglucamide solution, measuring the inactivated virus present in the final solution of step (d), and comparing the results of the final solutions of step (c) and step (e).

We claim: 
     
         1 . A method of purifying a biological product solution of interest having an unidentified enveloped virus contaminant comprising;
 (a) incubating the biological product solution of interest with an N-methylglucamide solution;   (b) inactivating any potential enveloped virus contaminant present in the biological product solution; and   (c) purifying the biological product solution.   
     
     
         2 . A method as recited in  claim 1 , wherein the enveloped virus comprises an RNA or DNA virus, single stranded or double stranded virus. 
     
     
         3 . A method as recited in  claim 1 , wherein an enveloped virus has been spiked into the biological product solution. 
     
     
         4 . A method as recited in  claim 2 , wherein the virus further comprises a retrovirus. 
     
     
         5 . A method as recited in  claim 1 , wherein the virus belongs within one or more of the following viral families selected from the group consisting of: Retroviridae, Flaviviridae, Togaviridae, Coronaviridae, Filoviridae, Rhabdoviridae, Bunyaviriae, Orthomyxoviridae, Paramyxoviridea, Arenaviridae, Hepadnaviridae, Herpesviridae, Baculoviridae and Poxviridae. 
     
     
         6 . A method as recited in  claim 1 , wherein the enveloped virus comprises a xenotropic leukemia virus, pseudorabies virus, or bovine viral diarrheal virus. 
     
     
         7 . A method as recited in  claim 1 , wherein the N-methylglucamide is composed of a fatty acid hydrophobic chain with carbon length of n (n≥3) linked by an amide bond to an acyclic, hydrophilic glucose sugar moiety. 
     
     
         8 . A method as recited in  claim 1 , wherein the N-methylglucamide is selected from the group consisting of Mega 8, Mega 9, Mega 10, Mega 11, and Mega 12. 
     
     
         9 . A method as recited in  claim 1 , wherein the biological product is selected from the group consisting of recombinant proteins, antibodies, cytokines, plasma proteins, vaccine solution, nucleic acid therapeutics and gene therapy products. 
     
     
         10 . A method as recited in  claim 1 , wherein the biological product was produced from a eukaryotic cell. 
     
     
         11 . A method as recited in  claim 10 , wherein the eukaryotic cell comprises a mammalian cell. 
     
     
         12 . A method as recited in  claim 1 , wherein the biological product was produced from a prokaryotic cell. 
     
     
         13 . A method as recited in  claim 7 , wherein the concentration of the N-methylglucamide solution is in a range of 0.5 to 10.0 times of critical micelle concentration (CMC). 
     
     
         14 . A method as recited in  claim 1 , wherein the incubating step is accomplished at low temperature. 
     
     
         15 . A method as recited in  claim 8 , wherein the incubating step takes place at a temperature range of 1 to 37 degrees Celsius. 
     
     
         16 . A method of inactivating an enveloped virus in an enveloped virus solution, comprising;
 (a) adding an N-methylglucamide solution to the enveloped virus solution, and   (b) inactivating the enveloped virus in the enveloped virus solution.   
     
     
         17 . A method as recited in  claim 15 , wherein the enveloped virus has been spiked into the biological product solution. 
     
     
         18 . A method of purifying a biological product solution of interest having an unidentified enveloped virus contaminant, comprising;
 (a) incubating a biological product solution of interest with a standard solution;   (b) inactivating any potential enveloped virus contaminant present in the biological product solution of step (a);   (c) measuring the inactivated viruses present in the final solution of step (b)   (d) incubating a separate biological product solution of interest with a N-methylglucamide solution;   (e) measuring the inactivated virus present in the final solution of step (d)   (f) comparing the results of the final solutions of step (c) and step (e).