Asparaginase antibodies were associated with HLA-DRB1*07:01 genotype, amino acid composition, and binding affinity. Anti-asparaginase IgG antibody status was available for 502 ALL patients. A higher incidence of asparaginase antibodies was detected for patients with (A) HLA-DRB1*07:01 (P = 1.4 × 10−5), (B) a glycine amino acid within HLA-DRB1 amino acid position 73 (P = 5.2 × 10−5), or (C) HLA-DRB1 alleles with predicted high-affinity binding for asparaginase (P = 1.4 × 10−3). Ala, alanine; Gly, glycine; het, heterozygous; homo, homozygous.