Development of a Chemiluminescent TSH Assay Using Magnetic Beads with monoclonal antibodies

Abstract

The assay of Thyroid stimulating hormone (TSH) levels helps in screening for hyperthyroidism and hypothyroidism. TSH assays have undergone improvements from the initial RIA methods to IMA methods after discovery of Monoclonal antibody technology with incorporation of chemiluminescent and fluorescent molecules further improving the sensitivity and automation of these assays.This project set out with the aim of using magnetic beads with monoclonal antibodies in order to establish and develop chemiluminescent TSH assay. Magnetic beads were prepared using 5 ml, 2.5% w/v particles. The particles were separated by use of a magnet and 0.25 ml acetate buffer 0.1 M, pH 5.0 added once the supernatant was aspirated. Using 50 µl of antibody concentration, (5 mg/ml) serial dilutions were made to give antibody concentrations of 0.5, 0.25 and 0.125 mg/ml respectively. The magnetic particles were added to a centrifuge tube and successive separation, aspiration of the supernatant and resuspension of the particles carried out with addition of relevant buffers and BSA to a final particle concentration of 1% W/V. Plates were prepared using ELISA (sandwich) protocol and Antigen- antibody reactions with varying antigen concentrations performed. Theexperimentwas able develop a good procedure for Chemiluminescent TSH assay using magnetic beads with monoclonal antibodies.

Keywords

  • Chemiluminescence TSH assays.   •Magnetic beads with Monoclonal antibodies.    •Varying antigen concentration.

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