Measurement of Autoantibodies to Gastric H+,K+-ATPase (ATP4A/B) Using a Luciferase Immunoprecipitation System (LIPS)

Methods Mol Biol. 2019:1901:113-131. doi: 10.1007/978-1-4939-8949-2_10.

Abstract

The Luciferase Immuno Precipitation System (LIPS) enables the detection of specific serum antibodies by immunoprecipitation of recombinant antigens tagged with a luciferase reporter. Here we describe LIPS assays for the quantification of autoantibodies to the H+, K+-ATPase A (ATP4A) and B (ATP4B) subunits, two serological markers of autoimmune atrophic gastritis and pernicious anemia. In particular, we will describe the expression of luciferase-tagged recombinant ATP4A and ATP4B, their immunoprecipitation with test sera, the recovery and washing of immune-complexes with a protein-A coated resin, and the quantification of autoantibodies by addition of a luciferase substrate and the measurement of the light output from captured luciferase-tagged antigens.

Keywords: ATP4A; ATP4B; Atrophic gastritis; Autoantibodies; Autoimmune gastritis; EIA; Enzyme immunoassay; Gastric proton pump H+,K+-ATPase; LIPS; Luciferase immunoprecipitation system.

Publication types

  • Research Support, N.I.H., Extramural
  • Research Support, Non-U.S. Gov't

MeSH terms

  • Antigens / metabolism
  • Autoantibodies / analysis*
  • Glycine / metabolism
  • H(+)-K(+)-Exchanging ATPase / immunology*
  • Humans
  • Immunoprecipitation / methods*
  • Luciferases / metabolism*
  • Protein Biosynthesis
  • Transcription, Genetic

Substances

  • Antigens
  • Autoantibodies
  • Luciferases
  • H(+)-K(+)-Exchanging ATPase
  • Glycine