Original Article:
Sodium caprylate wash during Protein A chromatography as an effective means for removing protease(s) responsible for target antibody fragmentation
Lixia Hu, et al.
Protein Expression and Purification, 2021, 186, 105907.
10.1016/j.pep.2021.105907
Residual CHO cell proteases may fragment recombinant proteins expressed in CHO, resulting in product-related low-molecular-weight (LMW) species. To maximize the removal of potential proteases that contribute to fragmentation, this study optimized Protein A chromatography by adding sodium caprylate (SC) to the wash buffer.
The table below demonstrates the effect of SC washing on eluate HCP levels, eluate quality, and step yield. Upon optimization, fragmentation of Protein A eluate happened to a much lesser degree as compared to that of eluate from unoptimized process. In addition, SDS-PAGE as shown demonstrates that fragmentation of protein A eluates with the lowest HCP levels, from runs containing 100 mM SC in wash2 buffer, was significantly minimized (lane 4, circled). Taken together, the data suggest that SC washing is an effective method for removing HCPs, including endogenous proteases responsible for target antibody fragmentation, during protein A chromatography.
Chemicals Related in the Paper:
| Catalog Number | Product Name | Structure | CAS Number | Price |
|---|---|---|---|---|
| ACM1984061 | Sodium caprylate |  |
1984-06-1 | Price |
