Open AccessA sandwich enzyme immunoassay for detecting and measuring alzheimer's disease‐associated proteins in human brain tissue
Author(s) -
Ghanbari Hossein A,
Ghanbari Hossein A,
Kozuk Terry,
Miller Barney E.,
Riesing Sue
Publication year - 1990
Publication title -
journal of clinical laboratory analysis
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.536
H-Index - 50
eISSN - 1098-2825
pISSN - 0887-8013
DOI - 10.1002/jcla.1860040308
Subject(s) - polyclonal antibodies , horseradish peroxidase , immunoassay , microbiology and biotechnology , antibody , monoclonal antibody , chemistry , antigen , monoclonal , biology , enzyme , immunology , biochemistry
A simple, fast, and reliable immunoassay has been developed to detect and measure Alzheimer's disease‐associated proteins (ADAPs) in human brain tissue. This assay, called ALZ‐EIA (brain), was developed for Abbott's quantum system, in which 1/4‐in. beads are used as solid phase. The beads were coated with polyclonal IgG purified from sera obtained from rabbits immunized with a highly enriched Alzheimer's disease brain protein fraction for ADAP. The antigen (ADAP) is effectively captured by the polyclonal IgG coated on the beads. ALZ‐50 (a mouse monoclonal IgM) is used as the detection antibody, and bound ALZ‐50 is subsequently quantified by a horseradish peroxidase‐linked goat antimouse IgM and appropriate substrate. The assay is linear up to 0.5 absorbance units (r = 0.9), reproducible (CV less than lo%), sensitive, and specific. With preformulated reagents and standard supplies, the assay is simple and rapid; 120 data points can readily be generated in about 4 hr.