Firsty, I found that fucosylation level of anti-A protein antibody is significantly lower than anti-B protein antibody with ELISA using lectin. (Coating: A or B protein, Blocking, Sera, Biotinylated AAL, HRP conjugated streptavidin)
And I wanna confirm this Data, but it is so difficult to confirm with Lectin blotting. So I have an idea that could instead lectin blotting, that is firstly I would purify fucosylated substance from serum by AAL agarose resin and check the antibody level of anti-A and anti-B using ELISA. (Coating: A or B protein, Blocking, fractions from AAL agarose chromatography, HRP conjugated anti-human IgG) So I could check if the antibody level of A is significantly lower than B.
Can anyone advise me whether the design is reliable?
Thanks so much.
Your description is so confusing for me. Are you looking for a fucosylated substance in serum which interfere with the antigen-antibody affinity? what about if the A or B antibodies are more or less glycosilated themselves?.
Apparently in your first assay you are not measuring antibody levels but detecting some fucosylation in antibodies. What was your objective, why a lectin was introduced at the assay?.
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