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

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Autoantibodies

Part of the book series: Methods in Molecular Biology ((MIMB,volume 1901))

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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Acknowledgments

E.L. and B.A. acknowledge support from University Sapienza 2013–2014. H.W.D. and J.M.W. acknowledge support from NIH grant R01 DK052068 (to H.W.D.). I.M., C.B., L.P., and V.L. acknowledge that their work was carried out within the framework of the “Ivascomar project, Cluster Tecnologico Nazionale Scienze della Vita ALISEI, Italian Ministry of Research” and with the support of the Associazione Italiana per la Ricerca sul Cancro (AIRC, bando 5 × 1000 N_12182). The authors wish to thank Carlo Lombardoni for his help in drawing Fig. 1.

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Author notes

  1. Edith Lahner and Ilaria Marzinotto contributed equally to this work.

Authors and Affiliations

  1. Medical-Surgical Department of Clinical Sciences and Translational Medicine, Sant’Andrea Hospital, Sapienza University of Rome, Rome, Italy

    Edith Lahner & Bruno Annibale

  2. Division of Genetics and Cell Biology, IRCCS San Raffaele Scientific Institute, Milan, Italy

    Ilaria Marzinotto & Vito Lampasona

  3. Beta Cell Biology Unit, Diabetes Research Institute, IRCCS San Raffaele Scientific Institute, Milan, Italy

    Cristina Brigatti & Lorenzo Piemonti

  4. Barbara Davis Center for Diabetes, University of Colorado Anschutz Medical Campus, Aurora, CO, USA

    Howard Davidson & Janet Wenzlau

Authors
  1. Edith Lahner
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  2. Ilaria Marzinotto
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  3. Cristina Brigatti
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  4. Howard Davidson
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  5. Janet Wenzlau
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  6. Lorenzo Piemonti
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  7. Bruno Annibale
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  8. Vito Lampasona
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Corresponding author

Correspondence to Vito Lampasona .

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Editors and Affiliations

  1. Department of Autoimmunology, Statens Serum Institut, Copenhagen, Denmark

    Gunnar Houen

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Lahner, E. et al. (2019). Measurement of Autoantibodies to Gastric H+,K+-ATPase (ATP4A/B) Using a Luciferase Immunoprecipitation System (LIPS). In: Houen, G. (eds) Autoantibodies. Methods in Molecular Biology, vol 1901. Humana Press, New York, NY. https://doi.org/10.1007/978-1-4939-8949-2_10

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  • DOI: https://doi.org/10.1007/978-1-4939-8949-2_10

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  • Publisher Name: Humana Press, New York, NY

  • Print ISBN: 978-1-4939-8948-5

  • Online ISBN: 978-1-4939-8949-2

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