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.

Measurement of Autoantibodies to Gastric H+,K+-ATPase (ATP4A/B) Using a Luciferase Immunoprecipitation System (LIPS) / Lahner, E; Marzinotto, I; Brigatti, C; Davidson, H; Wenzlau, J; Piemonti, L; Annibale, B; Lampasona, V. - (2019), pp. 113-132. [10.1007/978-1-4939-8949-2_10].

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

Lahner E
Primo
;
Annibale B;
2019

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.
2019
Autoantibodies. Methods and Protocols
9781493989485
9781493989492 eBook
ATP4A; ATP4B; atrophic gastritis; autoantibodies; autoimmune gastritis; EIA; enzyme immunoassay; gastric proton pump H+,K+-ATPase; LIPS; luciferase immunoprecipitation system
02 Pubblicazione su volume::02a Capitolo, Articolo o Contributo
Measurement of Autoantibodies to Gastric H+,K+-ATPase (ATP4A/B) Using a Luciferase Immunoprecipitation System (LIPS) / Lahner, E; Marzinotto, I; Brigatti, C; Davidson, H; Wenzlau, J; Piemonti, L; Annibale, B; Lampasona, V. - (2019), pp. 113-132. [10.1007/978-1-4939-8949-2_10].
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Utilizza questo identificativo per citare o creare un link a questo documento: https://hdl.handle.net/11573/1261593
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