Development of an ELISA for quantifying lysozyme in hen egg white.
An indirect enzyme-linked immunosorbent assay (ELISA) by inhibition was developed for quantifying lysozyme in hen egg white (HEW), a protein of value in not only the food and pharmaceutical industries but also for poultry research. Various experimental conditions (coating, antibodies dilutions, samples dilutions, preparations, blocking agents, and incubation times) were assayed to optimize this assay to the quantification of HEW in egg white samples. HEW samples were diluted 1:3000 to avoid matrix effects, possibly resulting from lysozyme interaction with other egg white proteins. Assay linearity for lysozyme ranged from 0.38 to 4.8 mug/mL, with intra- and interassay variations of 6.8% and 7.6%, respectively, and the lower detection limit was 0.264 mug/mL. We found that lysozyme concentrations in albumen from eggs laid by a hen cohort ranged from 2.2 to 4.5 mg/mL, thus underlining interhen variability. Overall, these data present an ELISA assay that is simple, quick, sensitive, accurate, and has been specifically designed to determine lysozyme concentrations in egg white samples.
Vidal
Mary-Laure
ML
Institut National de la Recherche Agronomique, Station de Recherches Avicoles, 37380 Nouzilly, France.
Gautron
Joël
J
Nys
Yves
Y
D000818:Animals; D000918:Antibody Specificity; D002645:Chickens; D004527:Egg Proteins / Q000032:analysis; D004529:Egg White / Q000032:analysis*; D004797:Enzyme-Linked Immunosorbent Assay / Q000379:methods*; D005260:Female; D009113:Muramidase / Q000032:analysis*; D015203:Reproducibility of Results; D012680:Sensitivity and Specificity
D004527:Egg Proteins; D009113:Muramidase