Gene/Protein
Disease
Symptom
Drug
Enzyme
Compound
Pivot Concepts:
Gene/Protein
Disease
Symptom
Drug
Enzyme
Compound
Target Concepts:
Gene/Protein
Disease
Symptom
Drug
Enzyme
Compound
Query: EC:1.13.12.5 (
aequorin
)
1,451
document(s) hit in 31,850,051 MEDLINE articles (0.00 seconds)
We established a simultaneous bioluminescent assay utilizing
aequorin
(Aq) and biotinylated firefly luciferase (b-Luc); furthermore, we developed a highly sensitive and rapid tandem bioluminescent immunoassay (BLIA) involving the Aq-labeled Fab fragment and b-Luc-streptavidin complex. Minimum detection limits of Aq and b-Luc were 9.4x10(-21) mol assay(-1) (blank + 3 S.D.) and 3.6x10(-19) mol assay(-1) (blank + 3 S.D.), respectively. Measurements of two luminescent proteins were completed in 4 s with a single assay medium. In this study, prostatic acid phosphatase (PAP) and prostate specific antigen (PSA), which served as analytes, were measured in the tandem BLIA. PAP and PSA were detected by the Aq-labeled anti-Dig Fab fragment and b-Luc-streptavidin complex, respectively. The measurable ranges of PAP and PSA were 0.04-100 and 0.2-200 ng mL(-1), respectively. This technique was also applied to the simultaneous measurement of PSA and
alpha-fetoprotein
(
AFP
). Measurable ranges of PSA and
AFP
were 0.2-200 and 1.95-1000 ng mL(-1), respectively. Levels of PAP and PSA or PSA and
AFP
in human serum could be accurately determined with the proposed BLIA. Satisfactory correlations were observed between results obtained from the proposed BLIA and those derived from commercial kits.
...
PMID:Highly sensitive and rapid tandem bioluminescent immunoassay using aequorin labeled Fab fragment and biotinylated firefly luciferase. 1738 17
The mutated recombinant
aequorin
with a reactive cysteine residue (Cys-
aequorin
) was highly purified and then conjugated with a maleimide-activated antibody without significant loss of luminescence activity. The conjugate ratio of Cys-
aequorin
to heavy chain of immunoglobulin G (IgG) was estimated to be 1:1. To test the bioluminescent immunoassay with
aequorin
-labeled antibody,
alpha-fetoprotein
(
AFP
), a serological marker of liver cancer, was used as a model analyte. The measurable range of
AFP
was 0.02 to 200 ng/ml with the coefficient of variation between 2.1 and 4.5%.
...
PMID:Recombinant aequorin with a reactive cysteine residue for conjugation with maleimide-activated antibody. 1841 74
We compare
aequorin
, alkaline phosphatase and horseradish peroxidase as reporters for luminescent immunoassays using
alpha-fetoprotein
(
AFP
) as a model analyte. Biotinylated
aequorin
was prepared from mutated
aequorin
containing a reactive cysteine residue by chemical conjugation. The measurable range of
AFP
using this biotinylated
aequorin
was 0.02-200 ng/ml, with a lower background level than the other biotinylated enzymes tested.
...
PMID:Comparison of luminescent immunoassays using biotinylated proteins of aequorin, alkaline phosphatase and horseradish peroxidase as reporters. 1906 Mar 86