LabRoots 2019 - Flow Through Polishing and The Use of Multi Mode Ligand Libraries to Improve Process

Multi-mode chromatography has come to prominence in recent years due to the general utility of multi-mode ligands for product capture and polishing. In particular, their application in flow-through polishing has significantly improved process economy. However, significant optimisation of binding and elution conditions is often required to achieve the desired level of purity. To overcome the deficiencies of individual multi-mode ligands, we have developed a diverse library of Multi-Mode Mimetic Ligands™ incorporating both ionic and hydrophobic groups, which offer significant flexibility in use. The Multi-Mode Mimetic Ligand™ Library is organised into a 12 x 8 array micro-column block and comprises a diverse array of ligands with a broad spread of charge, hydrophobic and hydrophilic functionalities. The adsorbents are produced using an agarose support matrix and incorporate triazine attachment chemistry for ease of synthesis and superior stability, Screening can be performed in high-throughput mode using robotic liquid handling systems or by use of conventional hand-held multi-channel pipettes. This approach allows for rapid screening, and identification of potential ligands for a specific target. Furthermore, adjustment of the starting protein fraction to suit individual ligands is not required as the diversity of the library enables the identification of binding ligands which are best-suited to the existing conditions and impurities. This approach has been evaluated in a variety of applications, including development of ligands for the removal of specific contaminants in a flow through mode. Learning Objectives: 1. Understand how you can use a diverse library of synthetic ligands to screen for adsorbents for your molecules that are difficult to purify using standard adsorbents. The screening library is fast becoming a must-have technology in the tool-box of a process development scientist. 2. A case study of using the Multi-Mode Mimetic Ligand™ Library to deliver a flow-through polishing step to remove FXI from human blood plasma.

About the Author

Buzz Lobbezoo

As the Product Support & Development (PSD) Manager for Astrea Bioseparations Ltd ('Astrea'), Buzz leads a small dedicated team providing full technical support and information for Astrea's products. Buzz joined Astrea in 2004 as a Research Scientist in the downstream purification team. In 2011 he became the Applications & Downstream Processing Specialist, and in 2016 was appointed the new role of PSD Manager. Buzz obtained his Biochemistry degree from University of Kent, then spent over 14 years in both the Research (University of Cambridge) and Biotech/Biopharmaceutical sectors (Adprotech, Metris and Celltech (now UCB)) before joining Astrea. He has significant product profiling and processing experience having worked on small-scale purification applications to large-scale manufacturing processes.

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