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Detection of differentially expressed Beta Amyloid fragments from Alzheimer's Patients by Mass Spectrometry
Author(s) -
Roth Steven Howard,
Bulman Amanda,
Thulasiraman Vanitha,
Berardini Mark,
Rusa Mariana
Publication year - 2010
Publication title -
the faseb journal
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.709
H-Index - 277
eISSN - 1530-6860
pISSN - 0892-6638
DOI - 10.1096/fasebj.24.1_supplement.lb158
Subject(s) - chemistry , alzheimer's disease , antibody , amyloid beta , microbiology and biotechnology , amyloid (mycology) , amyloid precursor protein , genetically modified mouse , chromatography , biochemistry , pathology , transgene , disease , peptide , medicine , biology , immunology , inorganic chemistry , gene
Study Objective A MALDI surface with covalently immobilized antibody specific to n‐terminus Beta Amyloid fragments was used to enrich specific fragments from biological samples for detection on a Bruker MALDI TOF/TOF instrument. Comparison of healthy and Alzheimer's disease diagnosed samples was evaluated by this method to determine if Beta Amyloid fragments are differentially expressed between the groups. Methods Human CSF pools from diagnosed Alzheimer's disease patients and healthy donors were treated with 9 M urea, 2% CHAPS, diluted in PBS, and applied in specific volume to a 2 mm antibody coated spot demarcated by a hydrophobic barrier. A similar treatment was used to prepare human brain cortex lysates from post mortem samples of Alzheimer's disease diagnosed and non‐Alzheimer's disease individuals. In a third set, brain samples from APP transgenic and wild type mice were prepared in the same fashion. After allowing samples to interact with the antibody surface, washes were done using PBS, PBS with Triton 100X, and 20 mM HEPES. Alpha‐cyano‐4‐hydroxycinnamic acid was applied to each spot after drying. All samples were read in linear mode in a MALDI TOF/TOF instrument. Results In each set of biological sample types evaluated, the Alzheimer's diagnosed group was found to express Beta Amyloid fragments, while none were detected in any of the control groups.