Generation of lectin plate-binding assays to assess plasma VWF glycan expression. (A-F) Lectin plate-binding assays were validated by assessing binding to a series of different VWF glycoforms generated by ex vivo treatment of purified pdVWF with specific exoglycosidases (summarized in schematic). After glycosidase digestions, residual glycan expression on VWF glycoforms were analyzed using SNA (A), MAL-II (B), WGA (C), RCA-I (D), ECA (E), and UEA-I (F). pdVWF was digested with α2-3 neuraminidase (purple bars), α2-3,6,8,9-neuraminidase (blue bars), α2-3,6,8,9-neuraminidase followed by β1-4 galactosidase (red bars), or PNGase F (green bars), respectively. All ELISAs were performed in triplicate, and results were expressed as a percentage of binding to untreated pdVWF. Results presented represent the mean values ± standard error of the mean. *P < .05; **P < .01; ***P < .001; Mann-Whitney U test. ns, not significant.