Development and application of an optical biosensor immunoassay for α-lactalbumin in bovine milk |
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| Authors: | Harvey E. Indyk |
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| Affiliation: | 1. Fonterra Co-operative Group Ltd, Cnr No. 1 Road & SH 26, Waitoa, New Zealand;2. R J Hill Laboratories Ltd, Clyde St, Hamilton, New Zealand;1. Department of Chemistry, Biotechnology and Bioprocess Engineering, Federal University of São João Del Rei, Ouro Branco, MG 36420-000, Brazil;2. Department of Veterinary, Federal University of Viçosa, Viçosa, MG 36570-000, Brazil;3. Department of Food Technology, Federal University of Viçosa, Viçosa, MG 36570-000, Brazil;4. Department of Animal Science, Federal University of Viçosa, Viçosa, MG 36570-000, Brazil;5. Department of Animal Science, Federal University of Bahia, Salvador, BA 40110-909, Brazil;1. State Key Laboratory of Animal Nutrition, Institute of Animal Science, Chinese Academy of Agricultural Sciences, Beijing 100193, China;2. Institute of Animal Science and Veterinary Medicine, Anhui Academy of Agricultural Sciences, Hefei 230031, China;1. State Key Laboratory of Food Science and Technology, Nanchang University, Nanchang 330047, China;2. Sino-German Joint Research Institute, Nanchang University, Nanchang 330047, China;3. Department of Pharmacy, Jiangxi University of Traditional Chinese Medicine, Nanchang 330004, China |
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| Abstract: | An automated, label-free biosensor-based immunoassay for α-lactalbumin in bovine milk utilising surface plasmon resonance (SPR) detection is described. α-Lactalbumin content was estimated from the specific interaction with an anti-bovine α-lactalbumin antibody immobilised on the sensor surface in a direct-binding assay format, although an alternative inhibition assay format is also described. Samples were prepared for analysis by direct dilution into buffer. Ligand selection and analysis conditions are defined, and non-specific binding considerations evaluated. Performance parameters include a working range of 10–1000 ng mL?1, a method detection limit of 0.12 mg mL?1 in milk, an overall instrumental reproducibility relative standard deviation (RSDR) of 5.71%, a mean inter-assay RSDR of 7.61% for an infant formula control sample and a surface stability of approximately 500 cycles. Accuracy was confirmed by comparison against an independent liquid chromatographic method. The technique was applied to the measurement of the α-lactalbumin content of consumer milk, colostrum, whey protein concentrates and infant formulae, the temporal change during early bovine lactation and a preliminary study of thermal denaturation. |
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