|
| Investigators | Sample type | Proteomics techniques | Outcome |
|
Immuno affinity MS
Advantages: Identify mutations on
known amyloid proteins.
Disadvantages: Only applicable to
known amyloid proteins; Require
antibody of high specificity;
Mutation may destroy antigenic
epitope, leading to reduction of
antibody binding; Amyloid proteins
may deposit in tissues leading to low
serum levels; Not as efficient as
DNA sequencing. | Tachibana et al. [29] | Serum | Immunoprecipitation, MALDI-TOF | Spectral pattern matching to genetic mutation |
| Sen et al. [44] | Serum, cerebrospinal fluid | On-line immunoaffinity capture, ESI-MS | Informations on relative abundance of protein isoforms, requires specific antibodies |
| Bergen et al. [28, 45] | Serum | Automated-online affinity purification, ESI-MS | Clinical assay to detect transthyretin mutations |
| Nepomuceno et al. [46] | Serum | Immunoprecipitation, Fourier-transform ion-cyclotron-resonance MS | Able to detect 91% of known transthyretin mutations due to high mass accuracy |
| Lavatelli et al. [47] | Serum | Immunopurification, SDS-PAGE or 2DE, MS | Can detect posttranslational modifications |
|
Antibody-free serum or tissue extract analysis
Advantages: Identify mutations on
known amyloid proteins; Measure
relative abundance of mutant to
wide type proteins.
Disadvantages: Only applicable to
known amyloid protein. | Kishikawa et al. [48] | Serum | Multidimensional liquid chromatography, ESI-MS | Applied to serum samples without requiring specific antibodies |
| da Costa et al. [49] | Serum | SDS-PAGE fractionation, MALDI-Fourier transform ion cyclotron resonance MS | Quantitation of mutant to wild type transthyretin |
| Ueda et al. [50] | Tissue and serum | HPLC separation, SELDI-TOF | Quantitation of mutant to wild type transthyretin within 3 hours |
| Lavatelli et al. [25] | Fine needle aspiration of abdominal adipose tissue | 2DE, PMF by MALDI-TOF | Identification of amyloidogenic proteins |
|
Direct extraction from FFPE samples
Advantages: Identify unknown
amyloid proteins in tissue sections
or fat aspirates.
Disadvantages: Protein extraction
requires days. | Murphy et al. [11] | FFPE | HPLC purification, N-terminal sequencing, PMF | Successful identification |
| Murphy et al. [51] | FFPE | HPLC, MS/MS de novo sequencing | Successful identification |
| | | | |
| | | | |
|
LMD with LC-MS/MS
Advantages: Capture specific regions
of interest: amyloid deposits from
FFPE sections; Identify unknown
amyloid proteins in tissue sections;
Protein extraction within hours.
Disadvantages: Requires specialist
LMD equipment and expertise. | Rodrigues et al. [52] | FFPE of immunoglobulin deposits in nerve tissue | LMD, LC-MS/MS | Successful identification |
| Vrana et al. [55] | FFPE samples of four amyloid types, 50 cases in training set, 41 cases in validation set | LMD, LC-MS/MS, algorithm to assign amyloid type according to highest number of spectra | >98% specificity and sensitivity for the 4 amyloid types examined |
| Klein et al. [53] | FFPE of various amyloid types in nerve tissue | LMD, LC-MS/MS | Successful identification of all 21 cases including transthyretin mutations |
| Sethi et al. [54] | FFPE of immunoglobulin renal deposits | LMD, LC-MS/MS | Successful identification |
|
MS Imaging
Advantages: Detecting amyloids in
situ, allowing correlation to
deposits; No extraction steps.
Disadvantages: Specialist MS
imaging software and expertise
required. | Stoeckli et al. [58] | Frozen tissue | MSI | Localization of amyloid β peptides in mouse brain sections |
| Seely and Caprioli [59] | One 100 year old amyloid FFPE sample | In-situ tryptic digest and imaging MALDI-MS/MS | Successful identification of serum amyloid A |
| | | | |
| | | | |
| | | | |
| | | | |
|
