Multimodal chromatography media (resin), e.g., the new Capto™ adhere ImpRes, has been proven very successful in the polishing steps of monoclonal antibodies (MAbs). Selection of buffer composition and salt could have impact on the performance of such purification steps. In the development and verification of these new chromatography media, a lot of experience has been gained. Some examples of buffer and salt considerations are:
• Loading buffers using buffer blends resulting in high conductivities (> 4 mS/cm) may result in high binding capacities. However, MAbs may leak from the column during washes and/or early in the gradient. If MAb is eluting during the wash step, you could use buffer with lower conductivity and/or avoid buffer blending’s including Tris.
• Addition of salt, e.g. 100 mM NaCl to the elution buffer may for some MAbs result in sharper peaks.
• Sample load using high salt concentration (e.g. 500 - 1000 mM NaCl) may improve binding capacity and could also result in improved HCP clearance.
See application note 29-0273-38, Polishing of monoclonal antibodies using Capto adhere ImpRes in bind and elute mode,to learn more about the performance of these new polishing media. (Click on the View PDF icon below.)
Read more about applications of new multimodal chromatography media in the poster, "Polishing of a Domain Antibody Using Capto MMC ImpRes".