The Role of Differential Proteomics in Amyloid Typing: The Experience of the Pavia Referral Center

  • Francesca Lavatelli
  • Francesca Brambilla
  • Andrea Di Fonzo
  • Giovanni Ferraro
  • Giovanni Palladini
  • Pierluigi Mauri
  • Giampaolo MerliniEmail author
Part of the Current Clinical Pathology book series (CCPATH)


Upon demonstration of the presence of amyloid deposits in tissues, disease typing is necessary for establishing a correct diagnosis, planning a therapeutic strategy, defining the prognosis, and programming genetic counseling. Recent years have witnessed the introduction in the clinical setting of a new resource for the characterization of tissue amyloid deposits: mass spectrometry-based proteomics. Proteomics, centered on the identification of the deposited proteins through mass spectrometry (MS), is being increasingly used by major centers worldwide and allows typing of known as well as novel forms of these diseases.

The Amyloid Center at IRCCS Policlinico San Matteo/University of Pavia, in collaboration with ITB-CNR in Milan, has developed comparative proteomic approaches that allow amyloid typing on subcutaneous abdominal fat and tissue biopsies, and can be employed as first-line or support diagnostic methods on Congo red-positive samples. In addition, the “amyloid proteome signature” may be used as a novel molecular strategy to support the diagnosis of amyloidosis.

This chapter describes the clinical approaches that are currently in use at the Amyloid Center in Pavia, and how they are used in conjunction with the other available typing resources, in particular electron microscopy.


Amyloid Diagnosis Amyloid typing Abdominal fat Organ biopsy Proteomics Mass spectrometry Protein extraction Proteolytic digestion Label-free proteomics comparison Protein sequences databases Computational analysis 


  1. 1.
    Sipe JD, Benson MD, Buxbaum JN, Ikeda SI, Merlini G, Saraiva MJ, Westermark P. Nomenclature 2014: amyloid fibril proteins and clinical classification of the amyloidosis. Amyloid. 2014;29:1–4.Google Scholar
  2. 2.
    Lachmann HJ, Booth DR, Booth SE, Bybee A, Gilbertson JA, Gillmore JD, Pepys MB, Hawkins PN. Misdiagnosis of hereditary amyloidosis as AL (primary) amyloidosis. N Engl J Med. 2002;346:1786–91.CrossRefPubMedGoogle Scholar
  3. 3.
    Satoskar AA, Burdge K, Cowden DJ, Nadasdy GM, Hebert LA, Nadasdy T. Typing of amyloidosis in renal biopsies: diagnostic pitfalls. Arch Pathol Lab Med. 2007;131:917–22.PubMedGoogle Scholar
  4. 4.
    Solomon A, Murphy CL, Westermark P. Unreliability of immunohistochemistry for typing amyloid deposits. Arch Pathol Lab Med. 2008;132:14.PubMedGoogle Scholar
  5. 5.
    Palladini G, Verga L, Corona S, Obici L, Morbini P, Lavatelli F, Donadei S, Sarais G, Roggeri L, Foli A, Russo P, Zenone Bragotti L, Paulli M, Magrini U, Merlini G. Diagnostic performance of immuno-electron microscopy of abdominal fat in systemic amyloidoses. Amyloid. 2010;17:59–60.Google Scholar
  6. 6.
    Foli A, Palladini G, Caporali R, Verga L, Morbini P, Obici L, Russo P, Sarais G, Donadei S, Montecucco C, Merlini G. Role of minor salivary gland biopsy in the diagnosis of systemic amyloidosis: results of a prospective study in 62 patients. Amyloid. 2011;18(S1):80–2.CrossRefPubMedGoogle Scholar
  7. 7.
    Brambilla F, Lavatelli F, Di Silvestre D, Valentini V, Rossi R, Palladini G, Obici L, Verga L, Mauri P, Merlini G. Reliable typing of systemic amyloidoses through proteomic analysis of subcutaneous adipose tissue. Blood. 2012;119(8):1844–7.CrossRefPubMedGoogle Scholar
  8. 8.
    Lavatelli F, Valentini V, Palladini G, Verga L, Russo P, Foli A, Obici L, Sarais G, Perfetti V, Casarini S, Merlini G. Mass spectrometry-based proteomics as a diagnostic tool when immunoelectron microscopy fails in typing amyloid deposits. Amyloid. 2011;18(S1):64–6.CrossRefPubMedGoogle Scholar
  9. 9.
    Brambilla F, Lavatelli F, Di Silvestre D, Valentini V, Palladini G, Merlini G, Mauri P. Shotgun protein profile of human adipose tissue and its changes in relation to systemic amyloidoses. J Proteome Res. 2013;12(12):5642–55.CrossRefPubMedGoogle Scholar
  10. 10.
    Lavatelli F, Perlman DH, Spencer B, Prokaeva T, McComb ME, Théberge R, Connors LH, Bellotti V, Seldin DC, Merlini G, Skinner M, Costello CE. Amyloidogenic and associated proteins in systemic amyloidosis proteome of adipose tissue. Mol Cell Proteomics. 2008;7(8):1570–83.PubMedCentralCrossRefPubMedGoogle Scholar
  11. 11.
    Valentini V, Lavatelli F, Obici L, Donadei S, Perlini S, Palladini G, Merlini G. Proteomic characterization of amyloid deposits in transthyretin amyloidosis associated with various mutations. Amyloid. 2011;18(S1):61–3.CrossRefPubMedGoogle Scholar
  12. 12.
    Park SK, Venable JD, Xu T, Yates 3rd JR. A quantitative analysis software tool for mass spectrometry-based proteomics. Nat Methods. 2008;5(4):319–22.PubMedCentralPubMedGoogle Scholar
  13. 13.
    Mauri PL, Dehò G. A proteomic MudPIT approach to the analysis of RNA degradosome composition in Escherichia coli. Methods Enzymol. 2008;447:99–117.CrossRefPubMedGoogle Scholar
  14. 14.
    Sethi S, Vrana JA, Theis JD, Leung N, Sethi A, Nasr SH, Fervenza FC, Cornell LD, Fidler ME, Dogan A. Laser microdissection and mass spectrometry-based proteomics aids the diagnosis and typing of renal amyloidosis. Kidney Int. 2012;82(2):226–34.PubMedCentralCrossRefPubMedGoogle Scholar
  15. 15.
    Vrana JA, Theis JD, Dasari S, Mereuta OM, Dispenzieri A, Zeldenrust SR, Gertz MA, Kurtin PJ, Grogg KL, Dogan A. Clinical diagnosis and typing of systemic amyloidosis in subcutaneous fat aspirates by mass spectrometry-based proteomics. Haematologica. 2014;99(7):1239–47.PubMedCentralCrossRefPubMedGoogle Scholar

Copyright information

© Springer International Publishing Switzerland 2015

Authors and Affiliations

  • Francesca Lavatelli
    • 1
  • Francesca Brambilla
    • 2
  • Andrea Di Fonzo
    • 1
  • Giovanni Ferraro
    • 1
  • Giovanni Palladini
    • 1
  • Pierluigi Mauri
    • 2
  • Giampaolo Merlini
    • 1
    Email author
  1. 1.Department of Molecular Medicine and Amyloidosis Research and Treatment CenterFoundation IRCCS Policlinico San Matteo and University of PaviaPaviaItaly
  2. 2.Department of Proteomics and MetabolomicsInstitute for Biomedical Technologies (ITB-CNR)SegrateItaly

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