Abstract
During acute inflammation, human α1-acid glycoprotein (AGP) is subject to marked changes in branching of its glycans, its degree of fucosylation and the expression of sialyl-Lewisx)(SLex) groups. To be able to study these changes in glycosylation in more detail, a procedure was developed to isolate the different glycoforms of AGP in milligram amounts by preparative affinity electrophoresis (AE) with a free lectin as fractionating agent. The method was applied to isolate differently fucosylated forms of AGP with the fucose-specific lectinAleuria aurantia (AAL). AGP was separated into one non-reactive (AO) and four reactive (A1-A4) fractions. It was found that, in particular, the highly fucosylated fractions A3 and A4 contained the inflammation-induced SLex groups of AGP. Analysis by crossed affinoimmunoelectrophoresis (CAIE) with concanavalin A (Con A) of these different glycoforms of AGP showed that the presence of tri-and/or tetraantennary glycans, instead of diantennary glycans, was associated with a higher degree of fucosylation. Identical results were obtained by subjecting Con A-fractionated forms of AGP to CAIE with AAL as the affinocomponent. It is expected that this method of preparative AE can generally be applied to other glycoproteins, which can be separated in different glycoforms by CAIE using lectins.
Published Version
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