Predicting the Origins of Anti-blood Group Antibody Specificity: A Case Study of the ABO A-and B-antigens Supplementary Material

Makeneni S, et al., 22;5:397, Front Immunol, 2014

This article describes unique insights into the affinity and specificity of a mAb raised against blood group A antigen using computational modeling and the BLI. Despite versatility of this carbohydrate-antibody interaction in human blood transfusion medicine, very little information is known about the factors contributing to their high specificity. The 3D models generated for blood group A and B trisaccharides in complex with the antibody variable region, scFv AC1001 were able to identify key residues and interactions such as hydrogen bonding that are contributing to higher affinity and specificity. The affinity measurements involving the scFv (immobilized on to amine reactive AR2G sensors) and the blood group A trisaccharide (conjugated to BSA) were obtained using a BLI (Octet RED96) system. The kinetic binding constants were determined assuming a 1:1 interaction. The BSA-LeX trisaccharide conjugate is used as the negative control.

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