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Protein Misfolding Diseases pp 123–153Cite as

Assays for Light Chain Amyloidosis Formation and Cytotoxicity

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Part of the book series: Methods in Molecular Biology ((MIMB,volume 1873))

Abstract

Common biophysical techniques like absorption and fluorescence spectroscopy, microscopy, and light scattering studies have been in use to investigate fibril assembly for a long time. However, there is sometimes a lack of consensus from the findings of an individual technique when compared in parallel with the other techniques. In this chapter, we aim to provide a concise compilation of techniques that can effectively be used to obtain a comprehensive representation of the structural, aggregation, and toxicity determinants in immunoglobulin light chain amyloidosis. We start by giving a brief introduction on amyloid assembly and the advantages of using simple and readily available techniques to study aggregation. After an overview on preparation of protein to set up parallel experiments, we provide a systematic description of the in vitro techniques used to study aggregation in AL protein. Additionally, we thoroughly discuss the steps needed in our experience during the individual experiments for better reproducibility and data analysis.

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Acknowledgments

The methods described in this chapter have been published for the most part in research articles, supported by the National Institutes of Health (R01 GM 071514), the Mayo Foundation, the Seidler Professorship and Dr. Morie Gertz, and the generous donations of amyloidosis patients and their families. Luis M. Blancas-Mejia and Pinaki Misra contributed equally to this work.

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Author notes

  1. Luis M. Blancas-Mejia and Pinaki Misra contributed equally to this work.

Authors and Affiliations

  1. Department of Biochemistry and Molecular Biology, Mayo Clinic, Rochester, MN, USA

    Luis M. Blancas-Mejia, Pinaki Misra, Christopher J. Dick, Keely R. Redhage, Shawna A. Cooper & Marina Ramirez-Alvarado

  2. Institute for Research in Biomedicine (IRB Barcelona), Barcelona, Spain

    Marta Marin-Argany

  3. Department of Immunology, Mayo Clinic, Rochester, MN, USA

    Marina Ramirez-Alvarado

Authors
  1. Luis M. Blancas-Mejia
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  2. Pinaki Misra
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  4. Marta Marin-Argany
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  5. Keely R. Redhage
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  6. Shawna A. Cooper
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  7. Marina Ramirez-Alvarado
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Correspondence to Marina Ramirez-Alvarado .

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Editors and Affiliations

  1. BioISI – Biosystems and Integrative Sciences Institute, Faculty of Sciences University of Lisbon, Lisbon, Portugal

    Cláudio M. Gomes

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Blancas-Mejia, L.M. et al. (2019). Assays for Light Chain Amyloidosis Formation and Cytotoxicity. In: Gomes, C. (eds) Protein Misfolding Diseases. Methods in Molecular Biology, vol 1873. Humana Press, New York, NY. https://doi.org/10.1007/978-1-4939-8820-4_8

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  • DOI: https://doi.org/10.1007/978-1-4939-8820-4_8

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