Development of biotin-streptavidin amplified peptide antibody-based ELISA for quantification of αS1-casein in goat’s milk

Alpha-s1-casein plays significant roles that determine the chemical, technological, and allergy properties of goat's milk. The well-known advantages of ELISA have made it becomes the gold method for the rapid detection of food biomolecules in diverse analytical settings. This study aims to develop a sandwich ELISA for quantification of αS1-casein in goat milk, utilizing peptide IgG-biotin as detection antibody. Under optimized condition, the calculated LOD and LOQ were 0.063 and 0.21 μg/mL, respectively. Cross-reactivity test showed that the ELISA was able to discriminate between αS1-casein and other proteins albeit their high homology (∼85%). The recoveries of αS1-casein in real milk samples were within the acceptable range of 80–120%. The %CV ( 0.9) indicated good reliability of ELISA in correlation with HPLC. The ELISA will substantially benefit several parties for the assessment of goat αS1-casein in milk and milk products, which include goat breeders, goat’s milk product manufacturers, diagnostic laboratories, and consumers. •Biotin-streptavidin antibody-based ELISA with high sensitivity against αS1-casein.•High specificity of ELISA was proven using other proteins in goat and cow milk.•Good reliability of ELISA (CV < 20%; r > 0.9) for milk samples application.