Peptide maps are particularly useful for comparison of different lots of a therapeutic protein and for use as the first dimension for inference of the protein’s amino acid sequential structure, degrees and sites of glycosylation, and extent of post-translational modification. After hydrolysis of the protein, the resulting peptides (ranging in size from a few to several hundred amino acid residues) are separated using HPLC and the resulting chromatogram (peptide map) is analyzed. Crucial to the protocol is the ability to separate the peptides with high retention time precision so that comparison of maps does not regularly require an identification step, such as mass spectrometry, cross-reaction with antibodies, or partial sequencing.
As demonstrated in the following application note, the ExpressLC system, with its characteristic high-accuracy gradient profile, is ideal for fast and repeatable analysis of peptide standards and samples. The precision of the Microfluidic Flow Control (MFC) system allows excellent retention time and peak area precision, which are both critical attributes to effective peptide map analysis.
Read the Application Note in its entirety.
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