Double-antibody based immunoassay for the detection of β-casein in bovine milk samples

Goldmag-based enzyme-linked immunosorbent assay for determination of α-lactalbumin in milk

“…In vitro detecting technology can be classified into three categories: immunological detection technology, the allergen analysis technology based on DNA, and instrumental analysis testing technology. Immunological methods were enzyme-linked immunosorbent assay (ELISA) (Fuller, Goodwin, & Morris, 2006;Zhou et al, 2013), rocket immunoelectrophoresis (Moen et al, 2005), and immunochromatographic strip tests (Puerta, Diez-Masa, & de Frutos, 2006;Puerta, Jaulmes, De Frutos, Diez-Masa, & Vidal-Madjar, 2002). Assays based on DNA analysis were mainly polymerase reaction (PCR) and fluorescence quantitative PCR (Galan, Brohee, Silva, van Hengel, & Chassaigne, 2011).…”

Magnetic bead and gold nanoparticle probes based immunoassay for β-casein detection in bovine milk samples

“…2H (b)). The limit of detection (LOD), calculated as the mean value of 10 blank samples plus 3 times standard deviations of the mean (Peng et al, 2008), was 4.8 ng mL À 1 , which was more than 700 times lower than that obtained from MBs-antibody-HRP based immunoassay , 6-7 times lower than those of microplate-antibody-HRP based assay (Zhou et al, 2013) and AuNPs aggregation-based assay . While the procedure of the double-probe based immunoassay is as simple as that of these methods.…”

“…Activated HRP (type B) kits and MBs (350 nm in diameter) were obtained from Tai Tianhe (Beijing, China) and Wa Wasaina (Wu Han, China), respectively. Anti-β-CN McAb and PcAb were produced in our previous study (Zhou et al, 2013). All reactions were carried out in 96-well polystyrene microtiter plates (Stripwell plate 2592, Costar, Changchun, China).…”

“…In order to avert the drawback of the 96-well-microplate based assay which requires more than 30 min for each antigen-antibody reaction step (Zhou et al, 2013), we used MBs as a solid support that only 10 min was needed for each antigen-antibody reaction step. We also used AuNPs as a bridge for loading more amounts of HRP molecules which increased the sensitivity of the assay.…”

“…Immunoassays based on antibody are sensitive and high throughput procedure for quantitative detection and also successfully applied to measurement of milk protein content, such as enzyme-linked immunosorbent assay (ELISA) for αs1-CN (Black and Reynolds, 1998) and β-CN (Song et al, 2011;Zhou et al, 2013), immunomagnetic beads-based immunoassay for β-CN , and nephelometric immunoassay for αs-CN and κ-CN (Collard-Bovy et al, 1991).…”