A one-step, competitive electrochemiluminescence-based immunoassay method for the quantification of a fully human anti-TNFα antibody in human serum

• Published in: Journal of pharmaceutical and biomedical analysis 2005-07, Vol.38 (4), p.703-708
• Main Authors: Horninger, D., Eirikis, E., Pendley, C., Giles-Komar, J., Davis, H.M., Miller, B.E.
• Format: Article
• Language: English
• Subjects: Analysis; Analytical, structural and metabolic biochemistry; Anti-TNFα; Biological and medical sciences; Competitive immunoassay; Electrochemiluminescence; Fundamental and applied biological sciences. Psychology; General pharmacology; Medical sciences; Monoclonal antibody; Pharmacology. Drug treatments
• ISSN: 0731-7085; 1873-264X
• DOI: 10.1016/j.jpba.2005.01.036

A quantitative, one-step, competitive electrochemiluminescence (ECL)-based immunoassay for the determination of a fully human, anti-TNFα monoclonal antibody in human serum has been developed. A biotinylated, mouse anti-variable region-specific antibody and a ruthenium-labeled anti-TNFα 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.