A lectin immunosensor technique for determination of alpha(1)-acid glycoprotein fucosylation.
The fucosylation of alpha(1)-acid glycoprotein (AGP), an acute-phase protein, is known to change in association with inflammatory diseases. Thus, fucosylation of AGP could be a potential diagnostic or prognostic marker. The change in fucosylation has previously been investigated using crossed affinoimmunoelectrophoresis, high-pH anion-exchange chromatography, and lectin ELISA. This study describes a surface plasmon resonance-based affinity biosensor assay for quantification of the fucosylation of AGP. Diluted EDTA plasma or serum was injected directly in a BIACORE 2000 biosensor. AGP was captured on the sensor surface using immobilized antibodies and a fucose-binding lectin from Aleuria aurentia was then used for the detection of fucosylation. The feature of the biosensor makes it possible to determine both the amount of bound AGP and the amount of bound lectin. Using a calibration curve it was possible to obtain a fucosylation ratio that was independent of AGP concentration. The assay was validated against a lectin ELISA and used to follow inflammation in patients with severe burns.
Liljeblad
M
M
Department of Biomedicine and Surgery, Linköping University, Linköping, SE-581 85, Sweden.
Rydén
I
I
Ohlson
S
S
Lundblad
A
A
Påhlsson
P
P
D015374:Biosensing Techniques; D004797:Enzyme-Linked Immunosorbent Assay; D005643:Fucose / Q000378:metabolism*; D037102:Lectins / Q000378:metabolism*; D009961:Orosomucoid / Q000302:isolation & purification / Q000378:metabolism*
D037102:Lectins; D009961:Orosomucoid; D005643:Fucose