Modified radioimmunoassay versus ELISA to quantify anti-acetylcholine receptor antibodies in a mouse model of myasthenia gravis

J Immunol Methods. 2024 Nov:534:113748. doi: 10.1016/j.jim.2024.113748. Epub 2024 Sep 4.

Abstract

In mouse models of myasthenia gravis (MG), anti-acetylcholine receptor (AChR) antibodies can be quantified to monitor disease progression and treatment response. In mice, enzyme-linked immunosorbent assay (ELISA) is the gold standard to quantify these antibodies. However, this method requires antigen purification, which is both time-consuming and expensive. In humans, radioimmunoassay (RIA)-which is more sensitive than ELISA-is commonly used to quantify AChR antibodies. At present, however, no commercial RIA kits are available to quantify these antibodies in mice. The aim of this study was to compare a modified commercial human RIA kit to two ELISA methods to detect AChR antibodies in an experimental autoimmune mouse model of MG (EAMG). C57BL/6 J mice were immunized with purified AChR from Tetronarce californica (T-AChR). Serum samples were analyzed by RIA and two ELISAs (T-AChR and purified mouse AChR peptide [m-AChR]). The modified RIA showed excellent sensitivity (84.1 %) and specificity (100 %) for the detection of AChR antibodies. RIA showed a good agreement with T-AChR ELISA (κ = 0.69) but only moderate agreement with m-AChR ELISA (κ = 0.49). These results demonstrate the feasibility of modifying a commercially-available RIA kit to quantify AChR antibodies in EAMG. The advantage of this technique is that it eliminates the need to develop the entire methodology in-house and reduces inter and intra-laboratory variability.

Keywords: Autoantibodies; EAMG; ELISA; Myasthenia gravis; RIA.

Publication types

  • Comparative Study

MeSH terms

  • Animals
  • Autoantibodies* / blood
  • Autoantibodies* / immunology
  • Disease Models, Animal
  • Enzyme-Linked Immunosorbent Assay* / methods
  • Female
  • Humans
  • Mice
  • Mice, Inbred C57BL*
  • Myasthenia Gravis / blood
  • Myasthenia Gravis / diagnosis
  • Myasthenia Gravis / immunology
  • Myasthenia Gravis, Autoimmune, Experimental / blood
  • Myasthenia Gravis, Autoimmune, Experimental / diagnosis
  • Myasthenia Gravis, Autoimmune, Experimental / immunology
  • Radioimmunoassay* / methods
  • Receptors, Cholinergic* / immunology
  • Sensitivity and Specificity

Substances

  • Receptors, Cholinergic
  • Autoantibodies