Lectin-enzyme immunoassay of transferrin sialovariants using immobilized antitransferrin and enzyme-labeled galactose-binding lectin from Ricinus communis.
A heterologous lectin-enzyme immunoassay is described. Microtiter plate wells were coated with affinity-purified antibodies to human transferrin. After incubation with transferrin sialovariants, prepared by limited neuraminidase treatment and separated with chromatofocusing, a lectin-enzyme-streptavidin complex was added. A good correlation was obtained between the number of terminal galactose groups on transferrin and the response in the lectin-enzyme immunoassay using Ricinus communis agglutinin as the galactose-binding lectin. The results indicate that characterization of glycosylation is possible with less than a microgram of the glycoprotein available, using lectin-enzyme immunoassays.
Pekelharing
J M
JM
Department of Clinical Chemistry, Stichting Samenwerking Delftse Ziekenhuizen, The Netherlands.
Vissers
P
P
Peters
H A
HA
Leijnse
B
B
D004797:Enzyme-Linked Immunosorbent Assay; D037161:Galectins; D014644:Genetic Variation; D006388:Hemagglutinins; D007124:Immunoenzyme Techniques; D007074:Immunoglobulin G; D009439:Neuraminidase; D012794:Sialic Acids / Q000032:analysis*; D014168:Transferrin / Q000032:analysis / Q000235:genetics*
D037161:Galectins; D006388:Hemagglutinins; D007074:Immunoglobulin G; D012794:Sialic Acids; D014168:Transferrin; D009439:Neuraminidase