Agilent Mass-based fraction collection of synthetic peptides using the Agilent 1100 Series purification system说明书用户手册.PDF

Mass-based fraction collection of synthetic peptides using the Agilent 1100 Series purification system Application Ralf Moritz Nicola O’Reilly Abstract Synthetic peptides are becoming more and more important as drug can- didates in the treatment of a variety of diseases. In order to keep pace with the growing number of newly synthesized peptides, peptide purifica- tion should not represent the bottleneck in the drug discovery process. Mass-based fraction collection is an efficient technique for the purifica- tion of compounds with well-known masses. In contrast to fraction trig- gering with less specific detectors, only the compound of interest is puri- fied in each run. Hence, it is not necessary to pick target compounds out of a series of redundant fractions that have been collected. This Applica- tion Note demonstrates mass-based purification of a series of synthetic peptides with the Agilent 1100 Series purification system. The modular set-up of the system, a reliable fully-automated delay volume calibration and a comprehensive software package provide a versatile platform for purification tasks up to 100 mL/min. Crude peptides were successfully purified by reverse phase HPLC from less than 1 kDa to more than 10 kDa, covering a pI range from 4 to 13. Introduction Today, solid phase peptide synthe- A sis is the method of choice for large-scale peptide production. In order to remove scavengers and reaction by-products a subsequent purification step is indispensable. UV based !! !! !!!!!! Usually preparative reverse phase HPLC is applied to purify crude peptides after synthesis. In such B an approach target peptides are UV-based fractionated and their M