Determination of glyoxylyl-peptide concentration using oxime chemistry and RP-HPLC analysis.
Glyoxylyl-peptides are useful peptide derivatives in the context of hydrazone, oxime or thiazolidine ligations. We describe a method for the determination of glyoxylyl-peptide concentration based on the reaction of the alpha-oxo aldehyde group with an excess of O-benzylhydroxylamine. The amount of O-benzylhydroxylamine necessary to convert the alpha-oxo aldehyde group into the corresponding O-benzyloxime was determined by RP-HPLC analysis and corresponded to the quantity of glyoxylyl-peptide used in the experiment. The method is rapid, sensitive, accurate and allows the automated analysis of several samples.
Garcia
Jean-Michel
JM
UMR CNRS 8161, Biological Institute of Lille, 1 rue du Pr Calmette, 59021 Lille, France.
Far
Samia
S
Diesis
Eric
E
Melnyk
Oleg
O
D001671:Biochemistry / Q000379:methods*; D002851:Chromatography, High Pressure Liquid / Q000379:methods*; D006038:Glyoxylates / Q000737:chemistry; D010091:Oximes / Q000737:chemistry; D010455:Peptides / Q000032:analysis*; D015203:Reproducibility of Results
D006038:Glyoxylates; D010091:Oximes; D010455:Peptides