A flow-injection continuous filtration approach for the automatic determination of monoclonal antibodies

✍ Scribed by J.M. Fernández-Romero; M.D.Luque de Castro

• Publisher: Elsevier Science
• Year: 1996
• Tongue: English
• Weight: 661 KB
• Volume: 331
• Category: Article
• ISSN: 0003-2670
• DOI: 10.1016/0003-2670(96)00232-2

✦ Synopsis

An unsegmented continuous manifold for development of a heterogeneous immunoassay has been developed in which a microfilter located prior to the detector retains the precipitate formed in the immunochemical reaction. The system used to check the performance of the dynamic approach is monoclonal mouse immunoglobulin G (IgG) as analyte and its corresponding antimouse immunoglobulin G labelled with peroxidase. The derivatizing reaction used to monitor the immunoreaction is based on the oxidation of 4-hydroxyphenylacetic acid with hydrogen peroxide in the presence of the peroxidase bound to the antimouse IgG, which was monitored fluorimetrically (A,,=3 15 nm and X,,=425 nm). The method for the determination of monoclonal mouse -1gG shows a linear range between 1 and 200 ng ml-' (3=0.9997, n=lO) with an RSD < 3% at 10 and 100 ng ml-' (n=33). The 3a detection limit is 0.1 ng ml-'. The approach should be generally applicable for the determination of monoclonal antibodies.