Primary structure chromatographic

Chloupek, R.C., Harris, R.)., Leonard, C. K., Keck, R.G., Keyt, B.A., Spellman, M.W., Jones, A. J.S., and Hancock, W. S. (1989) Study of the primary structure of recombinant tissue plasminogen activator by re-versed-phase high-performance liquid chromatographic tryptic mapping. J. Chromatogr. 463 375-396. 

Once a peptide map is characterized using online MS, the chromatographic profile alone can serve as a routine analytical tool to monitor the protein s primary structure and covalent modifications, and is often used for batch release or stability testing of biopharmaceuticals. However, whenever in-depth characterization of a protein is needed, such as that required for comparability studies or reference material characterization, the peptide map should be coupled with MS to ensure a thorough examination of all peptides in the map. 

In this section a variety of analytical separations reported in the literature are reviewed to show the wide structural diversity of eluite which can be separated by RPC and to assist the reader in becoming similar with the use of this fluid chromatographic technique. The descriptions are ar-ranged according to the matrix in which an analyte is found or the area of - h istry in which the samples are generally encountered. Thus theophylline, for example, is regarded as a nucleotide and, for the most part, its analysis in food samples is found with appropriate cross references. On the other hand, the separations of pharmaceuticals found in serum, urine, and pharmaceutical samples are cited separately. It is hoped that this method of classification may serve the purposes of those wh e analytical interests are incidental to their primary research pursuits. 

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