This application note describes the use of an Agilent HPLC-Chip system coupled to an Agilent Accurate-Mass Q-TOF LC/MS system for analysis of in-process samples of recombinant protein production. Combined with the powerful data processing capabilities of Agilent MassHunter and BioConfirm software, the superior mass accuracy of the HPLC-Chip LC/MS platform enabled identifi cation of impurities during the production process. This type of analysis aids in the assessment of impurities found in protein pharmaceutical at the early phases of product development.
The document details the use of an Agilent HPLC-Chip system coupled to an Agilent 6520 Accurate-Mass O-TOF LC/MS system for analyzing in-process samples of a recombinant protein named P128. The study aims to identify impurities during the production process of this anti-staphylococcal protein, which is under early development for human use against MRSA.
The analysis utilizes Agilent MassHunter Qualitative Analysis and BioConfirm software to deconvolute multiply charged ions and extract features from complex mixtures. The text compares the efficiency of the Large Molecular Feature Extractor (LMFE) algorithm against traditional maximum entropy deconvolution, demonstrating faster processing times and better identification of components in crude cell lysates compared to SDS-PAGE analysis.
The flow rate from the nano LC pump to the analytical column is 600 nL/min.