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Measurement of autoantibodies to gastric H+,K+-ATPase (ATP4A/B) using a luciferase immunoprecipitation system (LIPS)

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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.

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Lahner, E., Marzinotto, I., Brigatti, C., Davidson, H., Wenzlau, J., Piemonti, L., … Lampasona, V. (2019). Measurement of autoantibodies to gastric H+,K+-ATPase (ATP4A/B) using a luciferase immunoprecipitation system (LIPS). In Methods in Molecular Biology (Vol. 1901, pp. 113–131). Humana Press Inc. https://doi.org/10.1007/978-1-4939-8949-2_10

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