US Patent No. 10,654,886


Patent No. 10,654,886
Issue Date May 19, 2020
Title Method For Separation Of Monomeric Polypeptides From Aggregated Polypeptides
Inventorship Monika Baehner, Munich (DE)
Adelbert Grossmann, Eglfing (DE)
Stefan Hepbildikler, Penzberg (DE)
Assignee HOFFMANN-LA ROCHE INC., Little Falls, NJ (US)

Claim of US Patent No. 10,654,886

1. A method for removing aggregates from a solution with high aggregate content, comprising the steps of:a) applying a solution comprising the polypeptide in monomeric form and in aggregated form, wherein the aggregated form is 20% or more as determined by size exclusion chromatography of the total. polypeptide in the solution, to a mixed-mode Chromatography material. and recovering the polypeptide from the mixed-mode chromatography material, wherein the polypeptide recovered from the mixed-mode chromatography material has a reduced aggregate content; and
b) applying the solution obtained in the previous step to a weak cation exchange chromatography material and recovering the polypeptide therefrom and thereby producing the monomeric polypeptide,
wherein the solution applied to the mixed-mode chromatography material and/or the weak cation exchange chromatography material has a pH value at least 0.2 pH units and at most 2.5 pH units above the isoelectric point of the polypeptide.