Abstract
The introduction of high-performance liquid chromatography (HPLC) for the analysis and separation of peptides and proteins in the late 1970s offered unrivalled advantages in terms of speed, resolution, sensitivity, and recovery when used for the purification of low microgram levels of complex mixtures of peptides and proteins (1,2). However, the conventional HPLC columns of typically 4.6 mm id and operated at flow rates of 1 mL/min resulted in peak volumes of approx 1 mL or even larger. The resultant sample concentrations (μg/mL) were not ideally suited to subsequent manipulations, and at such low concentrations, losses caused by nonspecific adsorption on either the chromatographic support or associated equipment (e.g., syringes, sample vials, recovery vials) were commonplace (3,4). Attempts to reduce peak volumes by operating at lower flow rates were shown to be associated with poor recoveries of, in particular, hydrophobic proteins (5,6).
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Nice, E.C., Aguilar, MI. (2004). Micropreparative HPLC of Peptides and Proteins. In: Aguilar, MI. (eds) HPLC of Peptides and Proteins. Methods in Molecular Biology™, vol 251. Springer, Totowa, NJ. https://doi.org/10.1385/1-59259-742-4:165
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DOI: https://doi.org/10.1385/1-59259-742-4:165
Publisher Name: Springer, Totowa, NJ
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