Analysis of the Glycosylation Profile of Disease-Associated Water-Soluble Prion Protein Using Lectins

Figure 1

The binding of the PrP to the capturing antibodies was saturated. After sonication, an excess of total proteins from , , and plasma, calculated in order to achieve a saturated binding of PrP to the coated antibodies, was loaded into 96-well ELISA plates that were precoated with the anti-PrP capturing antibody. The figure shows similar absorbance values for PrP from , , and plasma indicating that equivalent amounts of PrP were captured and then identified by the antibodies. Since similar amounts of PrP were captured by 3F4 and 6H4 antibodies in the sandwich lectin-ELISA, this figure serves to support that the minor quantity of sugars detected on ws-PrP with respect to classical PrP is not due to the capturing of a minor amount of ws-PrP, but it rather reflects a difference in lectin binding. Data are means ± SD and are representative of at least three independent assays and three different preparations, performed in duplicate.