An effective double antibody sandwich ELISA (DAS-ELISA) method based on monoclonal (mAb) and chicken egg yolk IgY antibodies was developed to determine phosvitin (PV) content in therapeutic and functional products. Leghorn laying hens were immunized with purified PV to produce anti-PV IgY antibody in the egg yolk. High anti-PV IgY titer obtained from the egg yolks collected during 4-10 weeks of the immunization period contained approximately 6.2% of specific anti-PV IgY in total IgY. The PV detection range of the DAS-ELISA and biotinylated DAS-ELISA was 16.8-90 and 7.5-40 ng/mL, respectively. However, biotinylated DAS-ELISA was the better method for PV quantification in terms of accuracy and sensitivity. This highly efficient PV detection method may recuperate the performance of the existing protein assay methods as well as facilitate future research on PV bioactivities and applications.
Stratum corneum (SC) of the skin, which is the primary barrier for the body, is continuously exposed to external factors such as ultraviolet (UV) and blue light, which induce the production of reactive oxygen species (ROS), and aldehydes derived from lipid peroxidation. 1,2 ROS cause the oxidative modification of protein, DNA damage, inflammation, and aging of the skin. 3 Carbonylated proteins (CPs) are a type of oxidative modification of proteins, which reflects the oxidative stress caused by ROS induced by UV radiation and lipid peroxidation in SC. 4,5 CPs are produced by oxidative cleavage of proteins or by direct oxidation of amino acid side chains of lysine, arginine, proline, and threonine. 6 CP-induced deterioration of the skin's barrier functions promotes skin aging
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