3-(10'-Phenothiazinyl)propionic acid is a potent primary enhancer of peroxidase-induced chemiluminescence and its application in sensitive ELISA of methylglyoxal-modified low density lipoprotein.
Using a full factorial design the optimization of experimental conditions of enhanced chemiluminescence reaction (ECR) catalyzed by horseradish peroxidase (HRP) in the presence of 3-(10'-phenothiazinyl)propionic acid (PPA) as a primary enhancer was performed. The effect of concentrations of PPA, hydrogen peroxide, MORPH, luminol, and Tris on a ratio of peroxidase-catalyzed CL to background was studied. The detection limit value of HRP in ECR with PPA was 0.09 pM. Using PPA the ultra-sensitive chemiluminescent ELISA for determination of methylglyoxal-modified low density lipoprotein was developed. The detection limit value for the developed method was 0.5 ng mL(-1). The obtained results open up very promising perspectives for using PPA to improve the sensitivity of enzyme immunoassay kits.
Department of Chemistry, Lomonosov Moscow State University, 119991 Moscow, Russia. Electronic address: sakharovivan@gmail.com.
Ivan Yu
IY
Sakharov
Alexandra S
AS
Demiyanova
Anastasia V
AV
Gribas
Natalia A
NA
Uskova
Evgeny E
EE
Efremov
Marina M
MM
Vdovenko
D004797:Enzyme-Linked Immunosorbent Assay; D005163:Factor Analysis, Statistical; D006735:Horseradish Peroxidase / Q000737:chemistry; D006801:Humans; D006861:Hydrogen Peroxide / Q000737:chemistry; D057230:Limit of Detection; D008077:Lipoproteins, LDL / Q000097:blood*; D008163:Luminescent Measurements; D008165:Luminol / Q000737:chemistry; D009025:Morpholines / Q000737:chemistry; D010640:Phenothiazines / Q000737:chemistry; D011422:Propionates / Q000737:chemistry; D011765:Pyruvaldehyde / Q000737:chemistry*
D008077:Lipoproteins, LDL; D009025:Morpholines; D010640:Phenothiazines; D011422:Propionates; D008165:Luminol; D011765:Pyruvaldehyde; D006861:Hydrogen Peroxide; D006735:Horseradish Peroxidase
(CL); (CV); (ECR); (ELISA); (HRP); (LDL); (MG); (MORPH); (PPA); (RLU); (SPTZ); 3-(10′-Phenothiazinyl)-propinonic acid; 3-(10′-phenothiazinyl)-propane-1-sulfonate; 3-(10′-phenothiazinyl)propionic acid; 4-morpholinopyridine; Chemiluminescence; Enhancement; Enzyme immunoassay; Horseradish peroxidase; Methylglyoxal-modified low density lipoprotein; Peroxidase; chemiluminescence; coefficient of variation; enhanced chemuliminescence reaction; enzyme-linked immunosorbent assay; low density lipoprotein; methylglyoxal; relative luminescence units