title: Analytical Application of Flow Immunosensor in Detection of Thyroxine and Triiodothyronine in Serum.
abstract: In this study, an immunosensor based on kinetic exclusion analysis (KinExA) was used for thyroxine (T4) and triiodothyronine (T3) estimation. A KinExA™ 3200 instrument was used for this analysis, which is an automated flow fluorimeter designed to separate free unbound antibody binding sites in reaction mixtures of antibody, antigen, and antibody-antigen complex. A T3-BSA- and T4-BSA-coated polymethyl methacrylate (PMMA) bead microcolumn is generated inside the flow cell of the instrument. A sample mixture containing T3 and T4 with their respective monoclonal antibodies and their complexes are drawn past the microbead column. The unbound T3 or T4 monoclonal antibody binding sites are captured by their respective T3 and T4 antigens coated on the PMMA beads as bovine serum albumin conjugates. Fluorescently labeled secondary antibodies bind to the T3 or T4 antigen-antibody complex to generate fluorescence intensity for analysis. The limit of detection for the T3 and T4 assays was found to be 0.06 and 1.9 ng mL-1with acceptable precision values. The convenience of the automated KinExA format may be valuable in medical diagnostic laboratories.
authors: 1 Department of Pharmaceutical Chemistry, College of Pharmacy, King Saud University , Riyadh, Saudi Arabia .
authors: Tanveer A
authors: TA
authors: Wani
authors: 2 Department of Biochemistry, College of Science, King Saud University , Riyadh, Saudi Arabia .
authors: Seema
authors: S
authors: Zargar
authors: 3 Department of Genetics, King Faisal Specialist Hospital and Research Centre , Riyadh, Saudi Arabia .
authors: Salma
authors: S
authors: Majid
authors: 1 Department of Pharmaceutical Chemistry, College of Pharmacy, King Saud University , Riyadh, Saudi Arabia .
authors: Ibrahim A
authors: IA
authors: Darwish
mesh_terms: D015415:Biomarkers / Q000032:analysis / Q000097:blood; D002626:Chemistry, Pharmaceutical / Q000379:methods*; D005456:Fluorescent Dyes; D006801:Humans; D007118:Immunoassay / Q000379:methods; D013974:Thyroxine / Q000032:analysis / Q000097:blood*; D014284:Triiodothyronine / Q000032:analysis / Q000097:blood*
chemical_list: D015415:Biomarkers; D005456:Fluorescent Dyes; D014284:Triiodothyronine; D013974:Thyroxine
keywords: ELISA; immunosensor; thyroid disease; thyroxine; triiodothyronine