title: Development of an ELISA to detect circulating anti-asparaginase antibodies in dogs with lymphoid neoplasia treated with Escherichia coli l-asparaginase.
abstract: Resistance to Escherichia coli l-asparaginase in canine lymphoma occurs frequently with repeated administration, a phenomenon often attributed, without substantiation, to the induction of neutralizing antibodies. To test the hypothesis that treated dogs develop antibodies against the drug, we created an enzyme-linked immunosorbent assay (ELISA) to measure plasma anti-asparaginase immunoglobulin G responses. Using samples from dogs that had received multiple doses, specific reactivity against l-asparaginase was demonstrated, while naïve patients' samples were negative. The optimized ELISA appeared sensitive, with endpoint titers >1 600 000 in positive control dogs. Intra- and inter-assay coefficients of variation were 3.6 and 14.5%. The assay was supported by the observation that ELISA-positive plasma could immunoprecipitate asparaginase activity. When clinical patients were evaluated, 3/10 dogs developed titers after a single injection; with repeated administration, 4/7 dogs were positive. l-asparaginase antibodies showed reduced binding to the PEGylated drug formulation. The ELISA should prove useful in investigating the potential correlation of antibody responses with resistance.
authors.affiliation: Department of Clinical Sciences, North Carolina State University College of Veterinary Medicine, Raleigh, NC, USA.|Immunology Program, North Carolina State University College of Veterinary Medicine, Raleigh, NC, USA.
authors.forename: J A
authors.initials: JA
authors.lastname: Kidd
authors.affiliation: Department of Clinical Sciences, North Carolina State University College of Veterinary Medicine, Raleigh, NC, USA.|Immunology Program, North Carolina State University College of Veterinary Medicine, Raleigh, NC, USA.
authors.forename: P
authors.initials: P
authors.lastname: Ross
authors.affiliation: Department of Microbiology and Immunology, University of North Carolina at Chapel Hill School of Medicine, Chapel Hill, NC, USA.
authors.forename: A S
authors.initials: AS
authors.lastname: Buntzman
authors.affiliation: Department of Clinical Sciences, North Carolina State University College of Veterinary Medicine, Raleigh, NC, USA.|Immunology Program, North Carolina State University College of Veterinary Medicine, Raleigh, NC, USA.
authors.forename: P R
authors.initials: PR
authors.lastname: Hess
mesh_terms: D000818:Animals; D000906:Antibodies / Q000097:blood*; D000970:Antineoplastic Agents / Q000276:immunology / Q000627:therapeutic use*; D001215:Asparaginase / Q000276:immunology / Q000627:therapeutic use*; D004283:Dog Diseases / Q000097:blood / Q000188:drug therapy* / Q000276:immunology; D004285:Dogs; D004797:Enzyme-Linked Immunosorbent Assay / Q000662:veterinary; D004926:Escherichia coli / Q000201:enzymology*; D007074:Immunoglobulin G / Q000097:blood; D008223:Lymphoma / Q000097:blood / Q000188:drug therapy / Q000276:immunology / Q000662:veterinary*; D015203:Reproducibility of Results; D012680:Sensitivity and Specificity
chemical_list: D000906:Antibodies; D000970:Antineoplastic Agents; D007074:Immunoglobulin G; D001215:Asparaginase