Purpose: This study uses a high-throughput glycan microarray to evaluate the immunological evolution of antibodies to the glyco-antigen GD2. The goal is to determine germline and affinity mature antibody specificity and affinities/ Results: Affinity mature anti-GD2 antibodies 3F8 and ch14.18 had high affinity and were highly specific for the target GD2. Germline antibodies were also hihgly specific and had surprisingly high affinity. Conclusion: Antibodies to GD2 evolved from highly specific germlines. Highly specific germlines may be critical in evading autoimmunity issues.
Overall design
Germline variable regions of 3F8 and ch14.18 were determined. Germline and affinity mature antibody DNA were synthesized and transfected into HEK293 cell lines. Secreted antibodies were purified from supernatant using affinity chromatography. Antibodies were evaluated on multiple assay platforms, including glycan microarrays, protein microarrays, and cell binding assays.
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