This application note demonstrates peptide mapping performance for profiling of a tryptic digest of recombinant human erythropoietin (rhEPO) protein as a model digest for glycopeptides profiling. Like many protein therapeutics, rhEPO exhibits a great deal of heterogeneity due to modifications that occur during manufacturing. The AdvanceBio Peptide Mapping column, with an optimized C18 coating, provides excellent peptide selectivity and resolution across a broad elution range to enhance separations of the glycopeptides fragments for fast, efficient and sensitive mass spectrometry analysis.
This Technical Overview shows that the Agilent Buffer Advisor software together with the Agilent 1260 Infinity Bio-inert Quaternary LC System is ideal for automated protein separation by ionic strength gradients.
This Application Note describes an application solution for peptide mapping of a monoclonal antibody using the Agilent 1260 Infinity Bio-inert Quaternary LC System and proves the reliability of the solution. The bio-inertness and corrosion resistance of the instrument coupled with simple and reproducible methods make this solution particularly suitable for the QA/QC analysis of monoclonal antibody for the biopharmaceutical industry. The power range of the 1290 Infinity LC System (pressure versus flow) overcomes the challenge of increased back pressure caused by smaller particles and longer columns.
With the Agilent 1260 Infinity Bio-inert Quaternary LC System, phosphate compounds can be analyzed without any issues regarding the formation of phosphate-iron complexes found with stainless steel systems.