This document discusses the advantages of high-resolution analysis of intact antibodies, heavy chains, and light chains using ZORBAX Poroshell technology for high-velocity gradient separations. Method development is significantly improved with ZORBAX Poroshell HPLC columns, achieving up to 90% reduction in analysis time. ZORBAX Poroshell 300SB columns are available in various internal diameters and bonded phases for optimal sensitivity, resolution, and speed. The use of an HPLC instrument optimized for low-volume peaks of narrow width maximizes ZORBAX Poroshell resolution. The document also addresses the challenges in monoclonal antibody analysis.
This document provides a technical overview of optimizing the rapid High-Performance Liquid Chromatography (HPLC) analysis of monoclonal intact antibodies, light chains, and heavy chains using ZORBAX Poroshell columns. It highlights how high-velocity gradient separations with this technology result in high-resolution analysis with short run times and exceptional peak shapes. The text explains that method development is significantly more rapid, with analysis time reductions of up to 90% easily achieved compared to traditional wide-pore, totally-porous particle columns.
| Available internal diameters | 0.3, 0.5, 1.0, and 2.1-mm id |
|---|---|
| Available bonded phases | 300SB-C3, 300SB-C8, 300SB-C18, and 300Extend-C18 |
| Pore diameter | 300 A |
| Typical analysis time reduction | up to 90% |
| Flow rate comparison | 5 to 10 times faster |
Traditional wide-pore, totally-porous particle reversed-phase HPLC often results in unacceptably long analysis times and broad antibody peaks that compromise resolution.
The design features a solid silica core covered by a superficially porous surface, allowing large molecules to quickly diffuse in and out of the thin porous layer, enabling higher flow rates and short gradient times.
Available bonded phases include 300SB-C3, 300SB-C8, 300SB-C18, and 300Extend-C18.