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Activated Cross-linked Agarose for the Rapid Development of Affinity Chromatography Resins - Antibody Capture as a Case Study
Published on: August 16, 2019
Adsorption of chain type-specific ABO antibodies on Sepharose-linked A and B tetrasaccharides
Linda Lindberg1, Katharina Theinert, Jining Liu
1AbSorber AB, Department of Clinical Chemistry and Transfusion Medicine, Sahlgrenska Academy, University of Gothenburg, Sahlgrenska University Hospital, Gothenburg, Sweden.
Background:
Antigen-specific removal of anti-A and anti-B on immunoadsorption columns carrying the blood group A and B trisaccharides is one important component of some protocols used in ABO-incompatible organ transplantation. Because ABO antibodies exist requiring parts of the core saccharide chain for binding, the anti-A and -B-binding capacity of individual and combined, Sepharose-linked Types 1 through 4 A and B tetrasaccharides with that of the A and B trisaccharides was compared.
Study Design And Methods:
Sepharose-linked A and B tri- and tetrasaccharides were used to adsorb anti-A and -B from pooled blood group O serum. Remaining chain type-specific anti-A and -B were detected and quantified in enzyme-linked immunosorbent assays using wells coated with neoglycoproteins or recombinant mucins carrying A and B determinants on defined core saccharide chains.
Results:
Significantly more anti-A Type 3- and 4-specific immunoglobulin (Ig)G remained after adsorption on the A trisaccharide and the A Type 1 and A Type 2 tetrasaccharide than after adsorption on the A Types 3 and 4 tetrasaccharides. Selective adsorption of chain type-specific IgG anti-B was detected on Sepharose-linked B tetrasaccharides. In contrast, there were no chain type-specific IgM anti-A or -B. A combination of the A or B tetrasaccharides adsorbed a larger fraction of the IgG anti-A and -B repertoires than the corresponding trisaccharides.
Conclusion:
There are chain type-specific anti-A and anti-B IgG, and an adsorber based on a combination of Types 1 through 4 A or B tetrasaccharides will be a more efficient adsorber than an adsorber based on the A or B trisaccharides.
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