The sensitive and reproducible analysis of N-linked glycans of human immunoglobulin G is demonstrated, using an Agilent 1260 Infinity Bio-Inert Quaternary LC, with a novel HILIC column and fluorescence detection.
In this application note an Agilent ZORBAX Rapid Resolution High Definition (RRHD) 300-HILIC 1.8 ?m column and an Agilent AdvanceBio Peptide Mapping column are used in combination with time of flight (TOF) mass-spectrometry (MS) for mapping EPO protein. The work demonstrates the utility of HILIC as an orthogonal and complementary approach to reversed-phase LC/MS for peptide analysis.
To monitor monoclonal antibody titer and yield from cell-culture supernatants before expensive prep and large amounts of Protein A are employed, an analytical scale procedure helps determine the titer of monoclonal antibody for the optimal time for harvest of the mAb product. Here, pre-packed Agilent Bio-Monolith Protein A columns quickly capture mAb titer from cell supernatant.
In this work, an AdvanceBio Peptide Mapping column is used to generate a rapid and highly efficient peptide map at a traditional LC system pressure. The column achieves substantial improvements in peptide mapping during very fast run times and low system pressures, while still maintaining high peak-performance efficiency.
This Application Note shows method development for charge variant profiling of monoclonal antibodies using pH gradient elution with Agilent Buffer Advisor Software and online pH and conductivity monitoring.
In this study, hen egg white lysozyme was used as a model protein for PEGylation using N-terminal specific mPEG propionaldehyde (PEG aldehyde) in presence of cyanoborohydrate. A method for the analytical-scale purification of PEG lysozyme was developed using an Agilent 1260 Infinity Bio-Inert LC System and an Agilent Poroshell 120 SB-C18 column. Analytical-scale Fraction Collector with peak-based fraction collection was employed to collect the purified PEG lysozyme. The fractions were then re-analyzed by RP HPLC and SEC to demonstrate the homogeneity of the purified PEG conjugate.