Development and Application of Bovine β-lactoglobulin Gold Immunoassay Strip
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Abstract:
A colloidal gold immunochromatography method for the rapid determination of β-lactoglobulin (β-lg) in goat milk and its products were established. Bovine β-lg specific monoclonal antibody (monoclonal antibody, mAb) was prepared by hybridoma technique. Sodium citrate was used to reduce chloroauric acid. The 30 nm colloidal gold particles were formed and used to label bovine β-lgmAb. The competition method was used to develop immunochromatographic strips. A bovine β-lgmAb bag labeled with colloidal gold was placed on the gold label pad. Bovine β-lg and sheep anti-mouse IgG were labeled on nitrocellulose film as detection line (T line) and quality control line (C line), respectively. The optimal encapsulation concentration of bovine β-lg and secondary antibodies were both 1.0 mg/mL. The purity and titer of monoclonal antibody were all above 90% and above 10000 with high specificity. The limit of detection (LOD) of bovine β-lg was 3.13 μg/mL. There was no cross-reaction against other substrate components (bovine α-CN, β-CN, κ-CN, α-LA, BSA). The LOD of whole fat goat milk powder adulterated with skimmed cow milk powder was 0.5 %. The method was used to analyze the commercial goat milk and formula goat milk powder. The results of this method were consistent with that of the commercial ELISA kit. This method could be used to detect bovine β-lg in 5 min, and could be used to on-site rapid detection of sheep milk products.
