Optimization of oxidation of glycoproteins: an assay for predicting coupling to hydrazide chromatographic supports

A rapid, simple assay for aldehydes generated by oxidation of saccharide units in glycoproteins, using dyes containing hydrazide functionalities, is described. The assay is used, in conjunction with tests of biological activity, to predict oxidation conditions that will result in a maximum of active...

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Bibliographic Details
Published in:	Journal of chromatography Vol. 587; no. 2; p. 171
Main Authors:	Morehead, H W, Talmadge, K W, O'Shannessy, D J, Siebert, C J
Format:	Journal Article
Language:	English
Published:	Netherlands 20-12-1991
Subjects:	Aldehydes - analysis Aldehydes - metabolism Animals Cattle Chromatography - methods gamma-Globulins - analysis Glycoproteins - analysis Glycoproteins - metabolism Horseradish Peroxidase - analysis Humans Isoquinolines Oxidation-Reduction Serum Albumin, Bovine - analysis Xanthenes
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Summary:	A rapid, simple assay for aldehydes generated by oxidation of saccharide units in glycoproteins, using dyes containing hydrazide functionalities, is described. The assay is used, in conjunction with tests of biological activity, to predict oxidation conditions that will result in a maximum of active protein coupled to a hydrazide chromatographic support. Glycoproteins are labeled with Lucifer Yellow CH or Texas Red Hydrazide, and the extent of labeling is determined. Using the assay, it is shown that the efficiency of coupling to Affi-Prep Hydrazide is proportional to oxidation.
DOI:	10.1016/0021-9673(91)85153-7