Measurement of Autoantibodies to Gastric H plus ,K plus -ATPase (ATP4A/B) Using a Luciferase Immunoprecipitation System (LIPS)
Methods in molecular biology (Clifton, N.J.)(2019)
摘要
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.
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关键词
ATP4A,ATP4B,Atrophic gastritis,Autoantibodies,Autoimmune gastritis,EIA,Enzyme immunoassay,Gastric proton pump H+,K+-ATPase,LIPS,Luciferase immunoprecipitation system
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