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Quantification of small cyclic disulfide‐rich peptides
Author(s) -
Conibear Anne C.,
Daly Norelle L.,
Craik David J.
Publication year - 2012
Publication title -
peptide science
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.556
H-Index - 125
eISSN - 1097-0282
pISSN - 0006-3525
DOI - 10.1002/bip.22121
Subject(s) - chemistry , cyclic peptide , disulfide bond , absorbance , peptide , amino acid , gravimetric analysis , combinatorial chemistry , chromatography , organic chemistry , biochemistry
Cyclic disulfide‐rich peptides ranging in size from ∼14 to 29 amino acids have been found in a wide variety of organisms and have exciting biological and medicinal applications due to their stability and structure. Many of these peptides can be chemically synthesized, but their small size and limited number of chromophore‐containing amino acids make them difficult to quantify by methods routinely used for large proteins. A comparison of the precision and accuracy of gravimetric, UV‐ and NMR‐based methods in current use for the quantification of small peptides is presented for a representative set of cyclic disulfide‐rich peptides. The study shows that gravimetric and UV absorbance methods should be used with caution for small peptides and all methods should be carefully validated. For the routine quantification of small disulfide‐rich peptides, we recommend comparison of the analytical reverse‐phase high‐performance liquid chromatography trace or UV absorbance at 214 nm with that of a standard peptide solution quantified by amino acid analysis. An accurate quantification method that is simple and cost effective will assist in comparison of inhibition and activity data between different laboratories and peptides and correct calculation of synthesis yields. © 2012 Wiley Periodicals, Inc. Biopolymers (Pept Sci) 98: 518–524, 2012.