Multistep final filtration

Abstract

Herein is reported a method for the final filtration of concentrated polypeptide solutions comprising the combination of two immediately consecutive filtration steps with a first filter of 3.0 μm and 0.8 μm pore size and a second filter of 0.45 μm and 0.22 μm pore size.

1 . Method for producing an immunoglobulin solution comprising
 a) providing an immunoglobulin solution with a concentration of at least 100 g/l,   b) applying the immunoglobulin solution to a combination of a first and second filter unit, whereby the first filter unit comprises a pre-filter with a pore size of 3.0 μm and a main-filter with a pore size of 0.8 μm and the second filter unit comprises a pre-filter with a pore size of 0.45 μm and a main-filter with a pore size of 0.22 μm with a pressure of from 0.1 to 4.0 bar, and thereby producing an immunoglobulin solution.   
     
     
         2 . Method for producing an immunoglobulin comprising the following steps
 a) cultivating a cell, comprising a nucleic acid encoding the immunoglobulin   b) recovering the immunoglobulin from the cell or the cultivation medium,   c) purifying the immunoglobulin with one or more chromatography steps, and providing an immunoglobulin solution,   d) optionally adding a sugar, an amino acid and/or a detergent to the solution,   e) optionally concentrating the immunoglobulin solution to a concentration of 100 g/l or more with a method selected from diafiltration or tangential-flow filtration, and   f) applying the immunoglobulin solution of the previous step to a combination of a first and second filter unit, whereby the first filter unit comprises a pre-filter with a pore size of 3.0 μm and a main-filter with a pore size of 0.8 μm and the second filter unit comprises a pre-filter with a pore size of 0.45 μm and a main-filter with a pore size of 0.22 μm with a pressure of from 0.1 to 4.0 bar, and thereby producing an immunoglobulin.   
     
     
         3 . Method according to any one of the preceding claims, characterized in that the filter in the first and second filter unit have about the same filter area. 
     
     
         4 . Method according to any one of the preceding claims, characterized in that the immunoglobulin solution has a concentration of from 100 g/l to 300 g/l. 
     
     
         5 . Method according to any one of the preceding claims, characterized in that the immunoglobulin solution has a volume of from 3 liter to 100 liter. 
     
     
         6 . Method according to any one of the preceding claims, characterized in that the immunoglobulin is an anti-IL13 receptor alpha antibody or an anti-HER2 antibody. 
     
     
         7 . Method according to any one of  claims 2  to  6 , characterized in that the purifying is with a protein A affinity chromatography step and at least one step selected from cation exchange chromatography, anion exchange chromatography, and hydrophobic interaction chromatography. 
     
     
         8 . Method according to any one of the preceding claims, characterized in that the immunoglobulin solution has a concentration of 160 g/l or more and the applying to the combination of filters is by applying a pressure of 1.45 bar or more. 
     
     
         9 . Method according to any one of  claims 1  to  7 , characterized in that the immunoglobulin solution comprises a sugar and a surfactant and has a concentration of 125 mg/ml or more and the applying to the combination of the filter is by applying a pressure of 0.75 bar or less. 
     
     
         10 . Kit comprising a first filter with a pore size of from 3.0 μm to 0.8 μm and a second filter with a pore size of from 0.45 μm to 0.22 μm.