Optimizing Separation Conditions for the Isolation and Purification of Synthetic Peptides

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Thomas E. Wheat, Jo-Ann Jablonski, Elizabeth Gildea, and Jeffrey R. Mazzeo
ABRF The Association of Biomolecular Resource Facilities, March 31 - April 3, Tampa, Florida
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Peptides are used in basic research as well as in biopharmaceutical development. It is common to use synthetic peptides rather than isolating natural products. Such synthetic products may not be sufficiently homogeneous for use in experiments requiring bioactivity and specificity. Isolation and purification, therefore, become essential steps in a peptide synthesis facility. We have examined the factors that influence purification protocols. This includes selection of a column, operating parameters based on the properties of the peptides and the intended use of the product. A panel of synthetic peptides covering a range of properties including size, hydrophobicity, and isoelectric point is used for these evaluations.

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