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Second, the CRMS method was applied to an investigation of size variant impurity resolved by reversed phase - size exclusion 2D-LC. Finally, a separate size exclusion-reversed phase 2D-LC-CRMS method was developed to capture a wider range of process related impurities and reaction intermediates from the PEG-maleimide polymers throughout the conjugation process. The combination of these experiments using the 2D-LC-CRMS strategy enables sensitive characterization of the entire impurity profile of the high molecular weight multi-functional