Low ‐cost Customizable Microscale Toolkit for Rapid Screening and Purification of Therapeutic Proteins

This study demonstrates a versatile and customizable microscale column ( µCol) for biopharmaceutical separations using immobilized metal affinity chromatography (IMAC) as an example application to identify key parameters. µCols have excellent precision, efficiency, and reproducibility, can accommodate any affinity, ion‐exchange or size‐exclusion‐based resin and a re compatible with any HPLC system. µCols reduce reagent amounts, provide comparable purification performance and high‐throughput, and are easy to automate compared to current conventional resin columns. We provide a detailed description of the fabrication methods, resin packing methods, and µCo l validation experiments using a conventional HPLC system. Finite element modeling using COMSOL Multiphysics was employed to validate the experimental performance of the µCols. In this study, µCols were used for improving the purification achieved for granulocyte‐colony stimulating factor (GCSF) expressed using a cell‐free CHOin ‐vitro translation (IVT) system and were compared to a conventional 1 mL IMAC column. Experimental data revealed comparable purity with a 10 ‐fold reduction in the amount of buffer, resin and purification time for the μCols compared to conventional columns for similar protein yields.This article is protected by copyright. All rights reserved.
Source: Biotechnology and Bioengineering - Category: Biomedical Science Authors: Tags: ARTICLE Source Type: research