Abstract
Investigations of amyloidogenic diseases use synthetic peptides for cell-free and in vitro studies. However, amyloidogenic peptides often show intrinsic variability that markedly affects the reproducibility of experiments. Proof of physicochemical and biological variability with different batches of amyloidogenic peptides have been reported in literature. Here, we show that differences can be observed even within the same batch of Aβ1-42 peptide after storing lyophilised samples at -20°C. This change referred to as 'peptide aging' was reproduced with Aβ1-40 peptide samples by using a series of lyophilisation cycles, showing that lyophilisation, rather than preserving the physicochemical and biological features of Aβ peptides, introduces wide variability. To counteract synthetic peptide aging, we set up a procedure involving the sequential use of trifluoroacetic acid, formic acid and sodium hydroxide solutions that disaggregate preformed seeds and enriched Aβ peptide solutions into monomers and low-molecular-weight oligomers. This procedure enabled us to obtain reproducible physicochemical and biological features of Aβ peptides, irrespective of their age.
| Original language | English |
|---|---|
| Pages (from-to) | 71-80 |
| Number of pages | 10 |
| Journal | Amyloid |
| Volume | 16 |
| Issue number | 2 |
| DOIs | |
| Publication status | Published - 2009 |
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Keywords
- Aβ peptides
- Amyloid
- Lyophilisation
- Peptide aging reversal
- Trifluoroacetic acid
ASJC Scopus subject areas
- Internal Medicine
- Medicine(all)
Cite this
Overcoming synthetic Aβ peptide aging : A new approach to an age-old problem. / Manzoni, Claudia; Colombo, Laura; Messa, Massimo; Cagnotto, Alfredo; Cant, Laura; Del Favero, Elena; Salmona, Mario.
In: Amyloid, Vol. 16, No. 2, 2009, p. 71-80.Research output: Contribution to journal › Article
}
TY - JOUR
T1 - Overcoming synthetic Aβ peptide aging
T2 - A new approach to an age-old problem
AU - Manzoni, Claudia
AU - Colombo, Laura
AU - Messa, Massimo
AU - Cagnotto, Alfredo
AU - Cant, Laura
AU - Del Favero, Elena
AU - Salmona, Mario
PY - 2009
Y1 - 2009
N2 - Investigations of amyloidogenic diseases use synthetic peptides for cell-free and in vitro studies. However, amyloidogenic peptides often show intrinsic variability that markedly affects the reproducibility of experiments. Proof of physicochemical and biological variability with different batches of amyloidogenic peptides have been reported in literature. Here, we show that differences can be observed even within the same batch of Aβ1-42 peptide after storing lyophilised samples at -20°C. This change referred to as 'peptide aging' was reproduced with Aβ1-40 peptide samples by using a series of lyophilisation cycles, showing that lyophilisation, rather than preserving the physicochemical and biological features of Aβ peptides, introduces wide variability. To counteract synthetic peptide aging, we set up a procedure involving the sequential use of trifluoroacetic acid, formic acid and sodium hydroxide solutions that disaggregate preformed seeds and enriched Aβ peptide solutions into monomers and low-molecular-weight oligomers. This procedure enabled us to obtain reproducible physicochemical and biological features of Aβ peptides, irrespective of their age.
AB - Investigations of amyloidogenic diseases use synthetic peptides for cell-free and in vitro studies. However, amyloidogenic peptides often show intrinsic variability that markedly affects the reproducibility of experiments. Proof of physicochemical and biological variability with different batches of amyloidogenic peptides have been reported in literature. Here, we show that differences can be observed even within the same batch of Aβ1-42 peptide after storing lyophilised samples at -20°C. This change referred to as 'peptide aging' was reproduced with Aβ1-40 peptide samples by using a series of lyophilisation cycles, showing that lyophilisation, rather than preserving the physicochemical and biological features of Aβ peptides, introduces wide variability. To counteract synthetic peptide aging, we set up a procedure involving the sequential use of trifluoroacetic acid, formic acid and sodium hydroxide solutions that disaggregate preformed seeds and enriched Aβ peptide solutions into monomers and low-molecular-weight oligomers. This procedure enabled us to obtain reproducible physicochemical and biological features of Aβ peptides, irrespective of their age.
KW - Aβ peptides
KW - Amyloid
KW - Lyophilisation
KW - Peptide aging reversal
KW - Trifluoroacetic acid
UR - http://www.scopus.com/inward/record.url?scp=70849088649&partnerID=8YFLogxK
UR - http://www.scopus.com/inward/citedby.url?scp=70849088649&partnerID=8YFLogxK
U2 - 10.1080/13506120902879848
DO - 10.1080/13506120902879848
M3 - Article
C2 - 20536398
AN - SCOPUS:70849088649
VL - 16
SP - 71
EP - 80
JO - Amyloid : the international journal of experimental and clinical investigation : the official journal of the International Society of Amyloidosis
JF - Amyloid : the international journal of experimental and clinical investigation : the official journal of the International Society of Amyloidosis
SN - 1350-6129
IS - 2
ER -