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We evaluated the purification profiles of the following ceramic hydroxyapatite prepacked columns: 1.6 cm, 8 cm, 33 cm, and 45 cm inner diameter (ID) with 20 cm bed height. Each column used in the study was prepacked with CHT™ Ceramic Hydroxyapatite Type I Media (40 μm particle size). A sample mixture containing bovine serum albumin, lysozyme, and lactoferrin was loaded onto the column. A buffer of 5 mM sodium phosphate and 10 mM NaCl, pH 7.2, was used for equilibration. The sample mixture was then eluted with a linear gradient of 5 mM sodium phosphate, 10 mM NaCl, pH 7.2, to 400 mM sodium phosphate, pH 7.2.
We evaluated the purification profiles across all four columns to confirm the success of the method transfer and the scaled approach. The consistent purification profile results of the purification runs show that their peaks overlay from bench scale to the process-scale runs. This indicates that the Foresight Pro Column design and CHT Media packing methodology are optimized, delivering consistent results across the prepacked CHT Foresight Pro column sizes.
The use of traditional small-scale development techniques, combined with Foresight Pro prepacked columns, enables the research, process development, technical transfer, and manufacturing teams to efficiently achieve their objectives. This approach facilitates faster scaling techniques that require less material and minimizes risk. Prepacked chromatography columns offer several advantages including a reduction in buffers, resin usage, equipment footprints, labor costs, and increased flexibility in production schedule management.
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