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[82] Functional purification of synthetic proteins and protein fragments
Book chapter   Peer reviewed

[82] Functional purification of synthetic proteins and protein fragments

Methods in Enzymology, pp 631-638
1974
PMID: 4476047

Abstract

Purification of synthetic proteins and protein fragments by conventional separatory methods such as ion exchange chromatography, gel fractionation, or crystallization has often proved inadequate. This problem exists for solution syntheses, owing largely to the insolubility of large intermediate fragments, but especially for solid phase synthesis, because of the presence of unwanted failure sequences that may differ from the desired polypeptide by only subtle amino acid sequence variations. Inasmuch as purified synthetic proteins are of profound potential importance in the elucidation and engineering of conformational and functional features of proteins by planned chemical mutation of important residues, efforts have been made to develop efficient and specific purification techniques for such products. These techniques, categorized generally as functional purifications, are based on a property, or set of properties, exhibited by the correct sequence of the peptide or protein in question but not by at least the large majority of contaminant sequences. A specific and widely applied variety of functional purification is affinity chromatography, which involves separation on a solid support to which specific ligand is bound.

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