A rapid and sensitive immunoassay for tumor necrosis factor using magnetic monodisperse polymer particles

Abstract

A sensitive and rapid immunoassay for the detection of tumor necrosis factor (TNF) has been developed. Magnetic monodisperse polymer particles (M-280 Dynabeads) used as solid phase material, were coated with a neutralizing mouse monoclonal antibody to TNF. The coated Dynabeads were shown to have a more rapid binding capacity for recombinant (r) TNF as compared to standard immunowells coated with antibodies to TNF. The amount of TNF bound to the Dynabeads was quantified using either a polyclonal antibody to TNF or a mouse monoclonal antibody to TNF. The antibodies used for detection were either labelled with 125I or peroxidase. The linear assay range for the TNF standard curve was from 62 to 4000 pg/ml, and the assay time was less than 60 min. The sensitivity could be increased 5–8-fold by increasing the sample volume from 0.1 to 2 ml.