Enhanced chemiluminescent sandwich enzyme immunoassay for hen egg lysozyme.
A sensitive chemiluminescent sandwich-type enzyme immunoassay for hen egg lysozyme was developed. The assay was performed on polystyrene microtitre plates using immobilized specific polyclonal rabbit antibody against lysozyme, a peroxidase conjugate and the H2O2/luminol-enhanced chemiluminescence detection reagent. The chemiluminescent signal was detected using either a microplate luminometer, or photographic film in a camera luminometer. The detection limit for lysozyme was 0.3 ng/mL, and this was three times lower than that obtained using a colorimetric method with H2O2 and o-phenylendiamine as substrates. Recovery of the assay was 97-112% and the relative standard deviation ranged from 3.6% to 10.3%. The immunoassay overcame interference from the food sample matrix when lysozyme, used as a bacteriostatic agent, was measured.
Rauch
P
P
Department of Biochemistry and Microbiology, Institute of Chemical Technology, Prague, Czech Republic.
Poplstein
M
M
Hochel
I
I
Fukal
L
L
Ferri
E
E
Abagnato
C A
CA
Girotti
S
S
Roda
A
A
D000818:Animals; D002645:Chickens; D003124:Colorimetry / Q000379:methods; D004531:Eggs; D005260:Female; D005504:Food Analysis; D007124:Immunoenzyme Techniques; D007202:Indicators and Reagents; D008163:Luminescent Measurements; D009113:Muramidase / Q000032:analysis*; D012680:Sensitivity and Specificity
D007202:Indicators and Reagents; D009113:Muramidase