Abstract
It is well recognized that reverse-phase HPLC is one of the most powerful methods for peptide and protein isolation. The combination of high resolution and peak sensitivity with detection of the peptide bond at 200–220 nm has made this system an attractive complement to microsequence analysis. Although it is possible to perform reverse-phase HPLC using a variety of solvent systems, the trifluoroacetic acid/acetonitrile (TFA/MeCN) system in particular has become the most widely used because: (1) it has low absorbance at the 200–220 nm range; (2) the recovery of peptides is in the 90–100% range; (3) depending on the column and the protein, the recovery of proteins is also high; (4) peak shape and resolution are superior to the non-ion-pairing systems; and (5) the solvent is volatile and does not interfere with subsequent microsequence analysis.
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Shively, J.E. (1986). Reverse-Phase HPLC Isolation and Microsequence Analysis. In: Shively, J.E. (eds) Methods of Protein Microcharacterization. Biological Methods. Humana Press. https://doi.org/10.1007/978-1-59259-436-8_2
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DOI: https://doi.org/10.1007/978-1-59259-436-8_2
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