Protocol detail

Verified ABri, ADan, and Aβ Peptide Identification

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The identity of the ABri, ADan, and Aβ peptides in the LMPC isolated extracts from FBD, FDD, and CAA+ subjects was verified through LC–MS/MS analysis as previously described with few modifications (Michno et al., 2021 (link)). Briefly, the analysis with an alkaline mobile phase was carried out using a Q Exactive quadrupole–Orbitrap hybrid mass spectrometer equipped with a heated electrospray ionization source (HESI‐II) (Thermo Scientific) and UltiMate 3000 binary pump, column oven, and autosampler (Thermo Scientific). The Q Exactive was operated in data‐dependent mode. The resolution settings were 70.000 and target values were 1 × 10⁶ both for MS and MS/MS acquisitions. Acquisitions were performed with 1 μscan/acquisition. Precursor isolation width was 3 m/z units, and ions were fragmented using higher‐energy collision‐induced dissociation at a normalized collision energy of 25.

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Michno W., Koutarapu S., Camacho R., Toomey C., Stringer K., Minta K., Ge J., Jha D., Fernandez‐Rodriguez J., Brinkmalm G., Zetterberg H., Blennow K., Ryan N.S., Lashley T, & Hanrieder J. (2022). Chemical traits of cerebral amyloid angiopathy in familial British‐, Danish‐, and non‐Alzheimerʼs dementias. Journal of Neurochemistry, 163(3), 233-246.

Publication 2022

Q Exactive quadrupole–Orbitrap HESI‐II UltiMate 3000

Hybrid Ions Isolation Ms ms Peptides

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Variable analysis

independent variables

Alkaline mobile phase
Q Exactive quadrupole–Orbitrap hybrid mass spectrometer
Heated electrospray ionization source (HESI‐II)
UltiMate 3000 binary pump, column oven, and autosampler
Data‐dependent mode
Resolution settings of 70,000
Target values of 1 × 10^6 for both MS and MS/MS acquisitions
1 μscan/acquisition
Precursor isolation width of 3 m/z units
Higher‐energy collision‐induced dissociation at a normalized collision energy of 25

dependent variables

Identity of the ABri, ADan, and Aβ peptides in the LMPC isolated extracts from FBD, FDD, and CAA+ subjects

control variables

LC–MS/MS analysis as previously described with few modifications (Michno et al., 2021)

positive controls

Not explicitly mentioned

negative controls

Not explicitly mentioned

Annotations

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