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Reversed-Phase Selectivity for Peptide Mapping
Peptide mapping by reversed-phase LC-MS is a critical technique for confirming protein identity and assessing quality attributes in biopharmaceutical development. Selecting an optimal column is essential due to the diverse physicochemical properties of peptides. This study compares the performance of 13 reversed-phase columns using both a peptide standard mixture and a digested NIST monoclonal antibody. Key metrics such as peak capacity, selectivity, and peak asymmetry were evaluated across peptide subtypes including acidic, basic, and long-chain sequences. Results show core-shell columns, particularly those paired with bioinert hardware, offer enhanced resolution, efficiency, and reproducibility. Notably, PS C18 columns provided superior separation for acidic and basic peptides, while XB-C18 demonstrated strong performance across diverse peptide classes. These findings underscore the importance of tailored column selection for robust and comprehensive peptide mapping workflows.
Reversed-Phase Selectivity for Peptide Mapping
Peptide mapping by reversed-phase LC-MS is a critical technique for confirming protein identity and assessing quality attributes in biopharmaceutical development. Selecting an optimal column is essential due to the diverse physicochemical properties of peptides. This study compares the performance of 13 reversed-phase columns using both a peptide standard mixture and a digested NIST monoclonal antibody. Key metrics such as peak capacity, selectivity, and peak asymmetry were evaluated across peptide subtypes including acidic, basic, and long-chain sequences. Results show core-shell columns, particularly those paired with bioinert hardware, offer enhanced resolution, efficiency, and reproducibility. Notably, PS C18 columns provided superior separation for acidic and basic peptides, while XB-C18 demonstrated strong performance across diverse peptide classes. These findings underscore the importance of tailored column selection for robust and comprehensive peptide mapping workflows.
Document Type:
Technical Notes
Target Industries:
Techniques:
Brands:
Separation Modes:
Reversed Phase
Reversed-Phase Selectivity for Peptide Mapping
Peptide mapping by reversed-phase LC-MS is a critical technique for confirming protein identity and assessing quality attributes in biopharmaceutical development. Selecting an optimal column is essential due to the diverse physicochemical properties of peptides. This study compares the performance of 13 reversed-phase columns using both a peptide standard mixture and a digested NIST monoclonal antibody. Key metrics such as peak capacity, selectivity, and peak asymmetry were evaluated across peptide subtypes including acidic, basic, and long-chain sequences. Results show core-shell columns, particularly those paired with bioinert hardware, offer enhanced resolution, efficiency, and reproducibility. Notably, PS C18 columns provided superior separation for acidic and basic peptides, while XB-C18 demonstrated strong performance across diverse peptide classes. These findings underscore the importance of tailored column selection for robust and comprehensive peptide mapping workflows.
| Target Industries: | Biopharmaceutical Analysis |
| Techniques: | HPLC Reversed Phase |
| Brands: | Biozen LC, Kinetex, Luna Omega |
| Separation Modes: | Reversed Phase |
| Phases: | Biozen-Peptide-PS-C18, Biozen-Peptide-XB-C18, Kinetex Biphenyl, Kinetex C18, Kinetex C8, Kinetex F5, Kinetex Polar C18, Luna Omega C18, Luna Omega PS C18, Luna Omega Polar C18 |
Phenomenex is a technology leader committed to developing novel analytical chemistry solutions that solve the separation and purification challenges of researchers worldwide.
Phenomenex is a technology leader committed to developing novel analytical chemistry solutions that solve the separation and purification challenges of researchers worldwide.