Automated, High Productivity Purification and Analysis of Synthetic Peptides |
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Authors: | Thomas R Londo Terry Kehoe Steven A Kates Susan Hantman and Neal F Gordon |
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Institution: | (1) PerSeptive Biosystems, Inc., 500 Old Connecticut Path, Framingham, MA, 01721, U.S.A. |
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Abstract: | Methods for peptide assembly consist of techniques that allow for construction of complex sequences. The advantage of solid-phase methodologies is automation of the repetitive processes of deprotecting, washing, and coupling protected amino acids (acylation). However, for difficult sequences the crude product contains a variety of side products that must be removed to provide the desired target peptide in sufficient concentration and purity. This paper illustrates that high efficiency purification method-development can be achieved by combining purification and analysis on a single platform. Incorporation of fast LC-based assays using polystyrene-based POROS® Perfusion Chromatography media permitted rapid overall processing times from crude peptide purification through fraction pooling and product verification. Application of these technologies to the purification of peptides at scales of 100 mg is demonstrated. |
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Keywords: | analytical methods development high-performance liquid chromatography peptide purification process development process monitoring |
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