Charge Variant MS for Biotherapeutic Analysis with Sampling
Charge variant mass spectrometry is used for analyzing biotherapeutics with orthogonal and fractionated sampling.
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Imaged capillary isoelectric focusing (icIEF) is routinely performed for charge variant characterization of biotherapeutics but is incompatible with electrospray mass spectrometry. Advances in icIEF fractionation makes it possible to collect charge variant fractions offline for mass spectrometry characterization. How do users cope with the choice of a complex/ expensive linking of icIEF to Mass Spectrometry, or a tedious sample-bysample offline analysis?
Here we demonstrate an intelligent and automated approach to directly correlate charge profile from icIEF to MS data from the analysis of the collected fractions. Despite different LC and iCIEF separation mechanisms, the charge profiles from the two techniques can be superimposed and related to each other mathematically by ‘reconstructing’ the variant profile and displaying it on top of the LCMS profile. Direct comparison is highly desirable in any protein characterization environment because MS provides much deeper understandin of composition, but icIEF alone is ideal for routine.
