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:3.4.24.69 (
botulinum neurotoxin
)
1,901
document(s) hit in 31,850,051 MEDLINE articles (0.00 seconds)
Thrombin
and its aptamers have been well studied and widely used as models in aptamer based assays and sensors. Here we reported a
thrombin
-linked sandwich immunoassay for proteins to demonstrate new applications of
thrombin
and the aptamers, converting protein detection to analysis of
thrombin
label. In this assay, target protein was sandwiched by the capture antibody on a microplate and the biotinylated detection antibody.
Thrombin
bound to one biotinylated aptamer, and then the
thrombin
-labeled aptamer was attached on the sandwich complex through streptavidin-biotin interaction by using streptavidin as a linker.
Thrombin
catalyzed cleavage of fluorogenic peptide substrates, generating fluorescence signals for target detection. Among a few different anti-
thrombin
aptamers, the use of one nuclease resistant RNA aptamer having phosphorodithioate (PS2) modification on a specific backbone position enabled higher assay sensitivity due to its much higher affinity. This
thrombin
-linked sandwich immunoassay allowed detection of prostate-specific antigen (PSA) at 2 pM, an important protein related cancer disease, with high sensitivity and specificity. The strategy was general, and also enabled sensitive detection of
botulinum neurotoxin
type A (BoNTA) light chain, one toxin protein causing risk to human health. This assay combines advantages of antibody recognition, aptamer affinity labeling, high affinity of aptamers, and enzyme activity of
thrombin
. Labeling
thrombin
on the immunosandwich complex through simple affinity binding overcomes limitations of covalent conjugating enzyme on antibody in conventional immunoassay. This assay is promising in applications for protein detection.
...
PMID:A sensitive thrombin-linked sandwich immunoassay for protein targets using high affinity phosphorodithioate modified aptamer for thrombin labeling. 3159 65
