Column chromatography is a powerful separation tool for biopharmaceutical research and industry, with applications ranging from laboratory bench-scale purification to process development and commercial-scale manufacturing of biotherapeutics. Ensuring the highest quality of separations depends on many factors, including the technique used for packing the chromatography column. Well-packed columns provide for the best chromatographic separations. Successful column packing ensures proper mobile-phase distribution and resin contact. Scaling up to large columns can introduce many challenges. Manufacturers of chromatography media provide best-practice advice for packing their products. This eBook provides a starting point with key considerations for packing both traditional compressible and incompressible media.
Key factors affecting packing methodology differ by resin type. With compressible resins, the amount of media needed is determined by a compression factor, and additional compression needs to be applied during packing. Most such resins settle slowly, and once they are packed, there should be no headspace. Thus, rapid packing is not a significant factor. Most come in the form of a slurry, the amount of which equals the bed volume plus the compression factor. Low-shear impellers are suggested but not always required. Â Optimal packing requires precise measurement of settled (consolidated) bed height and compression factor.
With incompressible resins, the amount of resin needed is determined by tap-settled density, and bed consolidation is equivalent to bed packing. The media settle rapidly and require a headspace once packed. The best results are achieved with rapid packing. Many come in dry-powder form and require dispensing the correct amount followed by rehydration. Low-shear impellers are necessary to prevent particle fracturing and production of fines.Â Neither compression factor nor precise measurement of settled bed height are significant, but upflow bed conditioning should not be used during packing these media.
All these concerns are discussed in the eBook alongside some troubleshooting advice and links to sources for more information.
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