Chemiluminescent enzyme immunoassay of alpha-fetoprotein based on an adamantyl dioxetane phenyl phosphate substrate.
We have evaluated a new chemiluminescent substrate for the alkaline phosphatase (EC 3.1.3.1) label used in a Hybritech Tandem-E immunoassay of alpha-fetoprotein (AFP). The new substrate, adamantyl 1,2-dioxetane phenyl phosphate (AMPPD), emits light at 477 nm when acted upon by the enzyme. Detection limits for AFP with this method were 33 ng/L (mean of 20 replicates of the zero standard + 2 SD) and 470 ng/L (twice background). Between-batch CVs ranged from 4.31% to 9.60% for AFP in the range 29.1-132.0 micrograms/L. Comparison of results for 49 specimens assayed with use of the chemiluminescent kit and a colorimetric version of the AFP assay gave statistical values as follows: slope = 0.88, intercept = 4.19, and r = 0.94.
Thorpe
G H
GH
Department of Clinical Chemistry, Wolfson Research Laboratories, Queen Elizabeth Medical Centre, Birmingham, U.K.
Bronstein
I
I
Kricka
L J
LJ
Edwards
B
B
Voyta
J C
JC
D000218:Adamantane / Q000031:analogs & derivatives*; D000469:Alkaline Phosphatase; D000653:Amniotic Fluid / Q000032:analysis; D015415:Biomarkers / Q000032:analysis; D003124:Colorimetry; D005260:Female; D006801:Humans; D007124:Immunoenzyme Techniques; D008163:Luminescent Measurements; D011247:Pregnancy; D011933:Reagent Kits, Diagnostic; D013223:Statistics as Topic; D000509:alpha-Fetoproteins / Q000032:analysis* / Q000592:standards
D015415:Biomarkers; D011933:Reagent Kits, Diagnostic; D000509:alpha-Fetoproteins; C061523:3-(2'-spiroadamantane)-4-methoxy-4-(3''-phosphoryloxy)phenyl-1,2-dioxetane; D000469:Alkaline Phosphatase; D000218:Adamantane