A simple enzyme-linked immunosorbent assay (ELISA) for the neuron-specific gamma isozyme of human enolase (NSE) using monoclonal antibodies raised against synthetic peptides corresponding to isozyme sequence differences.
Monoclonal antibodies specific for the gamma isozyme of human enolase (known as neuron-specific enolase or NSE) have been raised against synthetic peptides after coupling to carrier protein: the selected peptides were those corresponding to regions of amino acid sequence difference between the alpha and gamma subunits of these closely similar isozymes. This technique gave monoclonal antibodies of high specificity and affinity. Two monoclonal antibodies raised against different peptides were used to develop a double-antibody sandwich enzyme-linked immunosorbent assay (ELISA), using one as the solid-phase antibody and the other conjugated to horseradish peroxidase to detect the bound NSE. This assay provides a simple and routine method of detecting NSE in serum samples from patients with small-cell carcinoma of the lung and related tumours.
Duncan
M E
ME
Department of Molecular and Cell Biology, University of Aberdeen, UK.
McAleese
S M
SM
Booth
N A
NA
Melvin
W T
WT
Fothergill
J E
JE
D000595:Amino Acid Sequence; D000911:Antibodies, Monoclonal; D000918:Antibody Specificity; D015153:Blotting, Western; D004797:Enzyme-Linked Immunosorbent Assay / Q000379:methods*; D006801:Humans; D007527:Isoenzymes / Q000032:analysis / Q000737:chemistry / Q000276:immunology; D008969:Molecular Sequence Data; D010455:Peptides / Q000276:immunology; D010751:Phosphopyruvate Hydratase / Q000032:analysis* / Q000737:chemistry / Q000276:immunology
D000911:Antibodies, Monoclonal; D007527:Isoenzymes; D010455:Peptides; D010751:Phosphopyruvate Hydratase