Back to results
US 2022/0026442 A1Published application
Method for using light scattering in real time to directly monitor and control impurity removal in purification processes
AssigneeAmgen Inc.
InventorsArthur C. HEWIG, III, Duke H. PHAN, Yinges YIGZAW, Robert BAILEY
Life of this patent
Estimating the term…
Abstract
The invention provides a method for controlling contaminants in biopharmaceutical purification processes by using light scattering and UV absorbance to establish a determinant. The invention makes use of multi-angle light scattering (MALS) and UV as a continuous monitoring system to provide information about the elution peak fractions in real-time instead of conventional pooling methods that rely on a predetermined percent UV peak max value to initiate the pooling process; regardless of product quality.
Claims5
1-41. (canceled)
A method for purifying a recombinant protein comprising:
- loading a mixture containing a recombinant protein and biological impurities on to a cation exchange chromatography resin;
- eluting by a salt gradient the recombinant protein from the cation exchange chromatography resin;
- measuring the intensity of light scattered by the eluate with a laser light scattering detector to obtain a light scattering (LS) signal;
- measuring the absorbance of the eluate with an ultraviolet (UV) absorbance detector to obtain an absorbance signal;
- calculating a ratio of the LS signal to the absorbance signal for each elution fraction to obtain a LS/UV fraction ratio;
- monitoring in real time during the elution of the recombinant protein the LS/UV fraction ratio over time for each elution fraction, wherein an increase in the LS/UV fraction ratio over time indicates an increase in the levels of biological impurities in the elution pool;
- terminating the elution when an increase in the LS/UV fraction ratio over time is detected;
- and
- recovering the recombinant protein from the eluate.
6 more elements
Dependencies are inferred from the claim text.