title: A one-step, competitive electrochemiluminescence-based immunoassay method for the quantification of a fully human anti-TNFalpha antibody in human serum.
abstract: A quantitative, one-step, competitive electrochemiluminescence (ECL)-based immunoassay for the determination of a fully human, anti-TNFalpha monoclonal antibody in human serum has been developed. A biotinylated, mouse anti-variable region-specific antibody and a ruthenium-labeled anti-TNFalpha antibody were the only specific reagents needed to develop the assay. A single incubation step of 2 h followed by ECL detection was used. The assay was capable of measuring the analyte in neat serum over approximately a 1600-fold range with higher concentrations measured following a single dilution. Assay accuracy, precision, and reproducibility were suitable to support pharmacokinetic studies of the analyte. This competitive assay format offers an alternative approach to the development of immunoassays for the measurement of macromolecules in complex matrices to support preclinical and clinical studies.
authors.affiliation: Department of Clinical Pharmacology and Experimental Medicine, Centocor, Inc., 145 King of Prussia Road, Radnor, PA 19087, USA.
authors.forename: D
authors.initials: D
authors.lastname: Horninger
authors.forename: E
authors.initials: E
authors.lastname: Eirikis
authors.forename: C
authors.initials: C
authors.lastname: Pendley
authors.forename: J
authors.initials: J
authors.lastname: Giles-Komar
authors.forename: H M
authors.initials: HM
authors.lastname: Davis
authors.forename: B E
authors.initials: BE
authors.lastname: Miller
mesh_terms: D000818:Animals; D000911:Antibodies, Monoclonal / Q000097:blood*; D000918:Antibody Specificity; D001172:Arthritis, Rheumatoid / Q000097:blood; D001710:Biotin; D006801:Humans; D007118:Immunoassay / Q000379:methods*; D007202:Indicators and Reagents; D008163:Luminescent Measurements; D051379:Mice; D012015:Reference Standards; D015203:Reproducibility of Results; D012428:Ruthenium; D014409:Tumor Necrosis Factor-alpha / Q000276:immunology*
chemical_list: D000911:Antibodies, Monoclonal; D007202:Indicators and Reagents; D014409:Tumor Necrosis Factor-alpha; D001710:Biotin; D012428:Ruthenium