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Improve Recovery and Reproducibility with Pre-Treatment of Analytical TSK-GEL G3000SWXL Size Exclusion Columns

les were analyzed by analytical SEC after BSA treatment, the HMW region was found to be enriched in host cell protein when compared to the HMW region of the same sample run on analytical SEC prior to BSA treatment. This suggests that the association between sample and column packing was most prevalent with the HCP as opposed to target molecule HMW species.




Conclusions:
Pre-treatment of SEC columns with BSA is a viable option to reduce secondary interactions and improve consistency of results between analyses. In this particular protocol, the dramatic improvement in the Improve Recovery and Reproducibility with Pre-Treatment of Analytical TSK-GEL G3000SWXL Size Exclusion Columns T.J. Higley, Tosoh Bioscience LLC and Richard Cornell, Wyeth BioPharma Figure 1. Overlay plot of the Protein A sample pre and post-treatment with BSA Sample % HMW pre treatment % HMW post treatment Post Protein A 1.1 3.4 Post Polishing chromatographic step1 1.5 1.5 Post Polishing chromatographic step 2 (final product) 0.2 0.2 Table 1: Recoveries of all three types of samples pre and post-treatment with BSA consistency of results associated with the protein A purified fraction, in contrast to the lack of effect seen on recovery of HMW species from downstream purification steps (chromatographic polishing steps 1 and 2), has provided the opportunity to further characterize the high molecular weight fractions of the protein A purified sample. This characterization has led to the conclusion that the results of SEC performed on samples from early purification steps may be negatively influenced by non-product related impurities derived from the cell c
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