Spot Analysis for Glycoprotein Determination in the Nanogram Range

Abstract
A flexible method of glycoprotein determination, with .mu.l-volumes, using spot analysis on cellulose acetate layers, is described. With [Aspergillus niger] glucose oxidase [EC 1.1.3.4] as an example of a glycoprotein and fluorescein isothiocyanate-labeled Concanavalin A, a sensitivity of 10 ng is reached; in combination with horseradish peroxidase [EC 1.11.1.7] 1 ng of glucose oxidase can be detected. A simultaneous determination of protein and glycoprotein with one single spot of 0.5, 1 or 2 .mu.l of a glycoprotein solution can be performed. The method is independent of many common external influences, e.g., dodecyl sulfate, Triton X-100, Nonidet P40, mercaptoethanol and desoxycholate. Only pretreatment of the glycoprotein with urea decreases the sensitivity.

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