Combined Method of Immunoaffinity Membrane Within Tubes and MALDI-TOF MS for Capturing and Analyzing Amyloid Beta

Amyloid beta 1-40 peptide was specifically isolated and analyzed from human plasma spiked with amyloid beta using a combined method of biotinylated anti-amyloid beta antibody binding to membrane-immobilized avidin (immunoaffinity membrane) and matrix-assisted laser desorption /ionization time-of-fli...

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Veröffentlicht in:Applied biochemistry and biotechnology 2015-12, Vol.177 (7), p.1565-1571
Hauptverfasser: Shimazaki, Youji, Takatsu, Yoko
Format: Artikel
Sprache:eng
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Zusammenfassung:Amyloid beta 1-40 peptide was specifically isolated and analyzed from human plasma spiked with amyloid beta using a combined method of biotinylated anti-amyloid beta antibody binding to membrane-immobilized avidin (immunoaffinity membrane) and matrix-assisted laser desorption /ionization time-of-flight mass spectrometry (MALDI-TOF MS). A solution of 10 μL containing 13.6 ng to 2.9 μg of amyloid beta peptide was examined in this method. After the isolated amyloid beta peptide from the spiked human plasma containing 2.9 μg of amyloid beta peptide was incubated in the presence of trifluoroacetic acid, fibrillization of the peptides was observed using a thioflavin T assay. Furthermore, an immunoaffinity membrane present on the inner wall of a tube (diameter 2 mm) captured the amyloid beta peptide from the spiked human plasma. Our results indicate that the combination of the immunoaffinity membrane procedure and MALDI-TOF MS can be used to capture and analyze the target antigens such as amyloid beta in micro-spaces.
ISSN:0273-2289
1559-0291
DOI:10.1007/s12010-015-1837-2