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Alpha-1 Antitrypsin Deficiency pp 223–226Cite as

Quantification of Z-AAT by a Z-Specific “Sandwich” ELISA

Part of the Methods in Molecular Biology book series (MIMB,volume 1639)

Abstract

This protocol describes an enzyme-linked immunosorbent assay (ELISA) to specifically detect Z-alpha-1 antitrypsin (AAT), the most common protein variant associated with alpha-1 antitrypsin deficiency. This “sandwich” ELISA relies on an anti-Z-AAT specific capture antibody and a HRP-conjugated anti-AAT detection antibody. This method would be of interest to identify and quantify Z-AAT in a variety of samples such as cell culture medium, cell or tissue lysate, animal or patient serum. Because this method is specific and sensitive, it would be particularly valuable for detection of Z-AAT in the presence of background M-AAT, for instance when quantifying silencing of Z-AAT in patients undergoing M-AAT augmentation therapy.

Key words

  • Alpha-1 antitrypsin
  • AAT
  • Z-AAT
  • Enzyme-linked immunosorbent assay
  • Sandwich ELISA
  • UMMS-PiZ-mAb

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References

  1. Wallmark A, Alm R, Eriksson S (1984) Monoclonal antibody specific for the mutant PiZ alpha 1-antitrypsin and its application in an ELISA procedure for identification of PiZ gene carriers. Proc Natl Acad Sci U S A 81(18):5690–5693

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  2. Janciauskiene S, Dominaitiene R, Sternby NH, Piitulainen E, Eriksson S (2002) Detection of circulating and endothelial cell polymers of Z and wild type alpha 1-antitrypsin by a monoclonal antibody. J Biol Chem 277(29):26540–26546. doi:10.1074/jbc.M203832200

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Acknowledgments

Development of the PiZ-specific antibody was funded by the Alpha-1 Foundation.

Author information

Authors and Affiliations

  1. Horae Gene Therapy Center, University of Massachusetts Medical School, Worcester, MA, USA

    Florie Borel, Qiushi Tang & Christian Mueller

  2. Department of Pediatrics, University of Massachusetts Medical School, Worcester, MA, USA

    Qiushi Tang & Christian Mueller

Authors
  1. Florie Borel
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  2. Qiushi Tang
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  3. Christian Mueller
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Corresponding author

Correspondence to Christian Mueller .

Editor information

Editors and Affiliations

  1. Horae Gene Therapy Center, University of Massachusetts Medical School, Worcester, Massachusetts, USA

    Florie Borel

  2. Horae Gene Therapy Center, University of Massachusetts Medical School, Worcester, Massachusetts, USA

    Christian Mueller

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Cite this protocol

Borel, F., Tang, Q., Mueller, C. (2017). Quantification of Z-AAT by a Z-Specific “Sandwich” ELISA. In: Borel, F., Mueller, C. (eds) Alpha-1 Antitrypsin Deficiency . Methods in Molecular Biology, vol 1639. Humana Press, New York, NY. https://doi.org/10.1007/978-1-4939-7163-3_22

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  • DOI: https://doi.org/10.1007/978-1-4939-7163-3_22

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  • Publisher Name: Humana Press, New York, NY

  • Print ISBN: 978-1-4939-7161-9

  • Online ISBN: 978-1-4939-7163-3

  • eBook Packages: Springer Protocols