Detection and quantitation of cellularly derived amyloid β peptides by immunoprecipitation-HPLC-MS

A quantitative method for detection of amyloid β peptides using immunoprecipitation-HPLC-mass spectrometry (IP-LC-MS) is described. Comparison of IP-LC-MS with sandwich ELISA revealed comparable results in the analysis of Aβ 1–40 and Aβ 1–42 derived from fetal guinea pig cell media and cell lysates....

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Veröffentlicht in:	FEBS letters 1998-07, Vol.430 (3), p.419-423
Hauptverfasser:	Clarke, Nigel J, Tomlinson, Andy J, Ohyagi, Yasumasa, Younkin, Steven, Naylor, Stephen
Format:	Artikel
Sprache:	eng
Schlagworte:	AcOH, acetic acid AD, Alzheimer's disease Alzheimer's disease Amyloid beta-Peptides - analysis Amyloid β Animals APP, amyloid precursor protein Aβ 1–40, amyloid β peptide consisting of APP residues 672–711 Aβ 1–42, amyloid β peptide consisting of APP residues 672–713 Aβ, amyloid beta peptide Brain Chemistry Cell Extracts CH3CN, acetonitrile Chromatography, High Pressure Liquid - methods Culture Media, Conditioned ELISA, enzyme linked immunosorbent assay Enzyme-Linked Immunosorbent Assay - methods HPLC HPLC, high performance liquid chromatography Humans Immunoprecipitation IP, immunoprecipitation LC, liquid chromatography Mass spectrometry Mass Spectrometry - methods MS, mass spectrometry n-PrOH, n-propanol PATRIC, position and time resolved ion counter PBS, phosphate buffered saline Peptides - analysis Precipitin Tests - methods RIPA buffer, 150 mM sodium chloride, 1% nonidet, 0.5% deoxycholate acid, 0.1% SDS, 50 mM Tris-base SDS, sodium dodecyl sulfate sELISA, sandwich ELISA Sensitivity and Specificity SIP, single ion profile Swine TFA, trifluoroacetic acid
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Zusammenfassung:	A quantitative method for detection of amyloid β peptides using immunoprecipitation-HPLC-mass spectrometry (IP-LC-MS) is described. Comparison of IP-LC-MS with sandwich ELISA revealed comparable results in the analysis of Aβ 1–40 and Aβ 1–42 derived from fetal guinea pig cell media and cell lysates. The use of IP-LC-MS not only allows a quantitative method for Aβ 1–40 and Aβ 1–42 peptides present in Alzheimer's disease (AD), but allows detection of other Aβ peptide species that may also play a role in the onset of AD in humans.
ISSN:	0014-5793 1873-3468
DOI:	10.1016/S0014-5793(98)00706-6