Process for Concentration of Antibodies and Therapeutic Products Thereof

Abstract

The present disclosure provides a process for concentrating proteins including an ultrafiltering, a diafiltering, and a second ultrafiltering sequence, at elevated temperatures, such as above about 30° C. The disclosure also includes a process for preparing highly concentrated antibody compositions, and highly concentrated antibody products.

1 . A process for preparing highly concentrated antibody compositions comprising: 
 a first ultrafiltering of a first antibody preparation to provide a second antibody preparation;    a diafiltering the second antibody preparation to provide a diafiltered intermediate antibody preparation; and    a second ultrafiltering of the diafiltered intermediate antibody preparation to provide a third antibody preparation;    wherein one or more of the first ultrafiltering, second ultrafiltering, and the diafiltering are accomplished at about 30° C. to about 50° C.    
   
   
       2 . The process of  claim 1  wherein one or more of the first ultrafiltering, the second ultrafiltering, and the diafiltering are accomplished at from about 35° C. to about 50° C.  
   
   
       3 . The process of  claim 1  wherein one or more of the first ultrafiltering, the second ultrafiltering, and the diafiltering are accomplished at from about 45° C.  
   
   
       4 . The process of  claim 1  wherein one or more of the first ultrafiltering, the second ultrafiltering, and the diafiltering are accomplished at 45° C. plus or minus 5° C.  
   
   
       5 . The process of  claim 1  wherein the first antibody preparation has an antibody concentration of from about 0.1 to about 10 grams per liter.  
   
   
       6 . The process of  claim 1  wherein the first antibody preparation has an antibody concentration of about 1 to about 5 grams per liter.  
   
   
       7 . The process of  claim 1  wherein the second antibody preparation has an antibody concentration of from about 10 to about 50 grams per liter.  
   
   
       8 . The process of  claim 1  wherein the second antibody preparation has an antibody concentration of about 20 to about 40 grams per liter.  
   
   
       9 . The process of  claim 1  wherein the third antibody preparation has an antibody concentration of from about 50 to about 250 grams per liter.  
   
   
       10 . The process of  claim 1  wherein the third antibody preparation has an antibody concentration of from about 100 to about 230 grams per liter.  
   
   
       11 . The process of  claim 1  wherein the third antibody preparation has an antibody concentration of from about 170 to about 200 grams per liter.  
   
   
       12 . The process of  claim 1  wherein the diafiltered intermediate antibody preparation and the third antibody preparation comprise the ultra-filter retentate.  
   
   
       13 . The process of  claim 1  wherein the intermediate antibody preparation has an antibody concentration of about 25 to about 35 grams per liter and the third antibody preparation has an antibody concentration of from about 170 to about 200 grams per liter.  
   
   
       14 . The process of  claim 1  wherein the antibody preparation comprises anti-IgE antibodies.  
   
   
       15 . The process of  claim 1  wherein the process is accomplished in from about 1 to 10 hours.  
   
   
       16 . The process of  claim 1  wherein the process is accomplished in from about 2 to 5 hours.  
   
   
       17 . The process of  claim 1  wherein the process is accomplished in about 3 hours.  
   
   
       18 . The process of  claim 1  wherein the first and the second ultrafiltering are accomplished with an ultra-filter membrane having a nominal pore size of about 5 to about 50 kilo Daltons.  
   
   
       19 . The process of  claim 1  wherein the first and the second ultrafiltering are accomplished with an ultra-filter membrane having a nominal pore size of about 10 to about 30 kilo Daltons.  
   
   
       20 . The process of  claim 1  wherein the first antibody preparation contains an antibody having an apparent molecular weight of about 100 to about 200 kilo Daltons.  
   
   
       21 . The process of  claim 1  wherein the first antibody preparation contains an antibody having an apparent molecular weight of about 150 kilo Daltons.  
   
   
       22 . The process of  claim 1  wherein the first ultrafiltering concentrates the first antibody preparation to provide the second antibody preparation having an antibody concentration of about 30 grams per liter and the second ultrafiltering concentrates the intermediate antibody preparation to provide the third antibody preparation having an antibody concentration of about 170 to about 200 grams per liter.  
   
   
       23 . The process of  claim 1  wherein the first ultrafiltering and the second ultrafiltering are accomplished with the same ultra-filter membrane.  
   
   
       24 . The process of  claim 1  wherein the first ultrafiltering and the second ultrafiltering are accomplished with a regenerated cellulose composite ultra-filter membrane.  
   
   
       25 . The process of  claim 1  wherein the diafiltering accomplishes a buffer exchange at constant volume, constant concentration, or both.  
   
   
       26 . The process of  claim 1  wherein the diafiltering accomplishes a buffer exchange of from about 5 to about 15 fold volumes.  
   
   
       27 . The process of  claim 1  wherein the diafiltering accomplishes a buffer exchange of from about 8 fold volumes.  
   
   
       28 . The process of  claim 1  wherein the diafiltering exchanges a first buffer for a second buffer.  
   
   
       29 . The process of  claim 28  wherein the first buffer comprises a mixture of aqueous sodium chloride and a TRIS buffer, and the second buffer comprises a mixture of aqueous histidine chloride and arginine chloride.  
   
   
       30 . The process of  claim 1  wherein the first ultrafiltering, the second ultrafiltering, and the diafiltering are accomplished with tangential flow filtration across an ultra-filter membrane.  
   
   
       31 . The process of  claim 1  wherein the first ultrafiltering, the second ultrafiltering, and the diafiltering are accomplished with tangential flow filtration across the same ultra-filter membrane.  
   
   
       32 . The process of  claim 1  wherein the yield of the third antibody preparation is greater than about 70 weight % based on the weight of antibodies in the first antibody preparation.  
   
   
       33 . The process of  claim 32  wherein the yield of the third antibody preparation is from about 80 to about 100 weight % based on the weight of antibodies in the first antibody preparation.  
   
   
       34 . The process of  claim 1  wherein the first ultrafiltering has a recirculation rate of from about 0.5 L/min/ft 2  to about 5 L/min/ft 2 .  
   
   
       35 . The process of  claim 1  wherein the ultrafiltering and diafiltering are accomplished at a transmembrane pressure of from about 10 to about 50 p.s.i.  
   
   
       36 . The process of  claim 1  wherein there is provided an antibody concentrate with a detectable bio-burden of less than about 100 CFU/mL.  
   
   
       37 . A process for concentrating proteins comprising: 
 a first ultrafiltering of a first protein mixture to provide a second protein mixture;    a diafiltering of the second protein mixture to provide a diafiltered protein mixture; and    a second ultrafiltering of the diafiltered protein mixture to provide a third protein mixture, wherein one or more of the first ultrafiltering, the diafiltering, and the second ultrafiltering are accomplished at from about 30° C. to about 50° C.