title: Development of ELISAs for quantification of HMFG1-specific human anti-mouse IgG and IgM antibodies.
abstract: The aim of this study was to develop and validate ELISAs for quantification of HAMA-IgM and HAMA-IgG in serum of patients with ovarian cancer who enrolled in a large international randomized phase III trial of intraperitoneal Yttrium-90-labeled HMFG1 murine monoclonal antibody therapy. The capture antibody of these 2 assays was the murine antibody HMFG1, while mouse anti-human IgM-HRP or mouse anti-human IgG(Fc)-HRP served as tracer antibodies. A pool of HAMA-positive serum samples was used to prepare a series of assay standards and another pool served as reference preparation. The analytical sensitivity of the HAMA-IgM assay was 2.5 arbitrary units per mL (AU/mL) and 4.7 AU/mL for the HAMA-IgG ELISA. Diluted serum samples showed good parallelism with the HAMA-IgM and HAMA-IgG standard dose-response curves. Within-assay coefficient of variation was 7.5% for HAMA-IgM and 6.5% for HAMA-IgG. Between-assay variation was 14.2% for HAMA-IgM and 15.3% for HAMA-IgG. The developed HAMA-IgM and HAMA-IgG ELISAs show satisfactory reliability criteria (sensitivity, parallelism and precision) and are suitable for monitoring of HAMA-IgM and HAMA-IgG responses in ovarian cancer patients. These ELISAs will be used to monitor the development of HAMAs in patients who received radioimmunotherapy with murine HMFG1.
authors.affiliation: Department of Obstetrics and Gynecology, Radboud University Nijmegen Medical Center, 6500 HB Nijmegen, The Netherlands. A.Oei@obgyn.umcn.nl
authors.forename: A L M
authors.initials: AL
authors.lastname: Oei
authors.forename: O C
authors.initials: OC
authors.lastname: Boerman
authors.forename: A
authors.initials: A
authors.lastname: Geurts-Moespot
authors.forename: J E
authors.initials: JE
authors.lastname: van Eerd
authors.forename: D
authors.initials: D
authors.lastname: van Tienoven
authors.forename: N
authors.initials: N
authors.lastname: Courtenay-Luck
authors.forename: C M G
authors.initials: CM
authors.lastname: Thomas
authors.forename: L F A G
authors.initials: LF
authors.lastname: Massuger
authors.forename: F C G J
authors.initials: FC
authors.lastname: Sweep
mesh_terms: D000818:Animals; D000888:Antibodies, Anti-Idiotypic / Q000276:immunology; D000911:Antibodies, Monoclonal / Q000276:immunology*; D014408:Biomarkers, Tumor / Q000097:blood*; D004797:Enzyme-Linked Immunosorbent Assay / Q000379:methods*; D005260:Female; D006801:Humans; D007141:Immunoglobulin Fc Fragments / Q000276:immunology; D007074:Immunoglobulin G / Q000097:blood* / Q000276:immunology; D007075:Immunoglobulin M / Q000097:blood* / Q000276:immunology; D000922:Immunotoxins / Q000276:immunology / Q000627:therapeutic use*; D007274:Injections, Intraperitoneal; D051379:Mice; D010051:Ovarian Neoplasms / Q000276:immunology* / Q000601:surgery / Q000628:therapy; D016499:Radioimmunotherapy / Q000379:methods; D012680:Sensitivity and Specificity; D015021:Yttrium Radioisotopes / Q000627:therapeutic use
chemical_list: D000888:Antibodies, Anti-Idiotypic; D000911:Antibodies, Monoclonal; D014408:Biomarkers, Tumor; D007141:Immunoglobulin Fc Fragments; D007074:Immunoglobulin G; D007075:Immunoglobulin M; D000922:Immunotoxins; D015021:Yttrium Radioisotopes; C469215:pemtumomab