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

FEBS Letters

Clarke, Nigel J¹  Younkin, Steven²  Tomlinson, Andy J¹  Ohyagi, Yasumasa³  Naylor, Stephen⁴ 

[1] Biomedical Mass Spectrometry Facility, Department of Biochemistry and Molecular Biology, Mayo Clinic, Rochester, MN 55905, USA;Department of Pharmacology, Mayo Clinic, Jacksonville, FL 32224, USA;Department of Neurology, Iizuka Hospital, 3-83 Yoshio-Machi, Iizuka City, Fukuoka 820, Japan;Biomedical Mass Spectrometry Facility, Department of Biochemistry and Molecular Biology, Department of Pharmacology and Clinical Pharmacology Unit, Mayo Clinic, Rochester, MN 55905, USA

关键词: Alzheimer's disease;    Amyloid β;    HPLC;    Immunoprecipitation;    Mass spectrometry;    AD;    Alzheimer's disease;    IP;    immunoprecipitation;    LC;    liquid chromatography;    HPLC;    high performance liquid chromatography;    MS;    mass spectrometry;    PATRIC;    position and time resolved ion counter;    APP;    amyloid precursor protein;    Aβ;    amyloid beta peptide;    Aβ 1–40;    amyloid β peptide consisting of APP residues 672–711;    Aβ 1–42;    amyloid β peptide consisting of APP residues 672–713;    SIP;    single ion profile;    ELISA;    enzyme linked immunosorbent assay;    sELISA;    sandwich ELISA;    PBS;    phosphate buffered saline;    SDS;    sodium dodecyl sulfate;    CH3CN;    acetonitrile;    TFA;    trifluoroacetic acid;    AcOH;    acetic acid;    n-PrOH;    n-propanol;    RIPA buffer;    150 mM sodium chloride;    1% nonidet;    0.5% deoxycholate acid;    0.1% SDS;    50 mM Tris-base;   

DOI  :  10.1016/S0014-5793(98)00706-6

学科分类：生物化学/生物物理

来源: John Wiley & Sons Ltd.

【 摘 要 】

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.

【 授权许可】

Unknown