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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.
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Document Type:
Technical Notes
Target Industries:
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.

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