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Amyloid-specific extraction using organic solvents

Kamiie, Junichi ; Aihara, Naoyuki ; Uchida, Yu ; Kobayashi, Daiki ; Yoshida, Yutaka ; Kuroda, Takeshi ; Sakaue, Motoharu ; Sugihara, Yutaka LU ; Rezeli, Melinda LU and Marko-Varga, György LU (2020) In MethodsX 7.
Abstract

Typing of amyloidosis by mass spectrometry (MS) based proteomic analysis contribute to the diagnosis of amyloidosis. For MS analysis, laser microdissection (LMD) is used for amyloid specific sampling. This study aimed to establish a method for selectively extracting amyloids from formalin-fixed, paraffin-embedded (FFPE) specimens by organic solvent instead of LMD. The extracts using dimethyl sulfoxide (DMSO), dimethylformamide (DMF), methanol, trifluoroethanol (TFE) or hexafluoro-2-propanol from FFPE brain of alzheimer's disease mouse model generated protein bands on SDS-PAGE, and Aβ was identified in the extract of DMF using mass spectrometry. The extract using DMSO from the kidney of a AA amyloidosis patient produced a protein band in... (More)

Typing of amyloidosis by mass spectrometry (MS) based proteomic analysis contribute to the diagnosis of amyloidosis. For MS analysis, laser microdissection (LMD) is used for amyloid specific sampling. This study aimed to establish a method for selectively extracting amyloids from formalin-fixed, paraffin-embedded (FFPE) specimens by organic solvent instead of LMD. The extracts using dimethyl sulfoxide (DMSO), dimethylformamide (DMF), methanol, trifluoroethanol (TFE) or hexafluoro-2-propanol from FFPE brain of alzheimer's disease mouse model generated protein bands on SDS-PAGE, and Aβ was identified in the extract of DMF using mass spectrometry. The extract using DMSO from the kidney of a AA amyloidosis patient produced a protein band in SDS-PAGE. This protein band was identified to be serum amyloid A (SAA) by Western blotting and mass spectrometry. Circular dichroism spectrometry revealed that the secondary structures of Aβ and transthyretin were converted to α-helices from β-sheets in TFE. Our results suggest that organic solvents can extract amyloids from FFPE specimens by converting their secondary structure. This method could eliminate the LMD step and simplified amyloid typing by MS analysis. • DMSO, DMF, methanol, TFE and HFIP can extract Aβ specifically from the FFPE brain of a Alzheimer’ disease mouse model. • DMSO can extract SAA specifically from a FFPE section of AA amyloidosis. • Secondary structures of Aβ and transthyretin converted from β-sheet to α-helix in TFE.

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organization
publishing date
type
Contribution to journal
publication status
published
subject
keywords
Amyloid, Amyloid-specific extraction from FFPE specimens using organic solvents, Formalin-fixed, Organic solvents, Paraffin-embedded, Protein extraction
in
MethodsX
volume
7
article number
100770
publisher
Elsevier
external identifiers
  • pmid:32140436
  • scopus:85078020933
ISSN
2215-0161
DOI
10.1016/j.mex.2019.100770
language
English
LU publication?
yes
id
bf5de93b-5d17-4689-b5bb-548c492227e9
date added to LUP
2021-01-11 13:51:58
date last changed
2021-04-06 01:34:17
@article{bf5de93b-5d17-4689-b5bb-548c492227e9,
  abstract     = {<p>Typing of amyloidosis by mass spectrometry (MS) based proteomic analysis contribute to the diagnosis of amyloidosis. For MS analysis, laser microdissection (LMD) is used for amyloid specific sampling. This study aimed to establish a method for selectively extracting amyloids from formalin-fixed, paraffin-embedded (FFPE) specimens by organic solvent instead of LMD. The extracts using dimethyl sulfoxide (DMSO), dimethylformamide (DMF), methanol, trifluoroethanol (TFE) or hexafluoro-2-propanol from FFPE brain of alzheimer's disease mouse model generated protein bands on SDS-PAGE, and Aβ was identified in the extract of DMF using mass spectrometry. The extract using DMSO from the kidney of a AA amyloidosis patient produced a protein band in SDS-PAGE. This protein band was identified to be serum amyloid A (SAA) by Western blotting and mass spectrometry. Circular dichroism spectrometry revealed that the secondary structures of Aβ and transthyretin were converted to α-helices from β-sheets in TFE. Our results suggest that organic solvents can extract amyloids from FFPE specimens by converting their secondary structure. This method could eliminate the LMD step and simplified amyloid typing by MS analysis. • DMSO, DMF, methanol, TFE and HFIP can extract Aβ specifically from the FFPE brain of a Alzheimer’ disease mouse model. • DMSO can extract SAA specifically from a FFPE section of AA amyloidosis. • Secondary structures of Aβ and transthyretin converted from β-sheet to α-helix in TFE.</p>},
  author       = {Kamiie, Junichi and Aihara, Naoyuki and Uchida, Yu and Kobayashi, Daiki and Yoshida, Yutaka and Kuroda, Takeshi and Sakaue, Motoharu and Sugihara, Yutaka and Rezeli, Melinda and Marko-Varga, György},
  issn         = {2215-0161},
  language     = {eng},
  publisher    = {Elsevier},
  series       = {MethodsX},
  title        = {Amyloid-specific extraction using organic solvents},
  url          = {http://dx.doi.org/10.1016/j.mex.2019.100770},
  doi          = {10.1016/j.mex.2019.100770},
  volume       = {7},
  year         = {2020},
}