Abstract: the separation of biologics, pharmaceutical industry uses commercial size exclusion high performance (SEC-HPLC) columns to assess, the major peaks, aggregates and degradants present in the product.
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The webinar will include developing a de novo method to eliminate the interactions of proteins or peptide with the residual silanols and improve the separation of proteins, peptides and impurities. This approach mitigates the problems associated with recovery of the monomeric and high molecular species for these biomolecule, which are retarded or bound on the column during separation.
Many conditions and additives were evaluated, high ionic strength, organic solvent and ion pairing agents. Several SEC-HPLC columns and mobile phases were evaluated for the SEC-HPLC assay of proteins and peptides to further diminish interactions with these residual silanols. Mobile phase optimization was evaluated based on plates, resolution, symmetry, recovery, RT closer to the real molecular weight  and purity and impurity recovery
Examples from peptide and antibodies are presented here with an identification of a novel ingredient in the mobile phase that reduced the interaction with full recovery of the monomeric and the aggregate species.