An Enzyme-Linked Immunosorbent Assay for Detection of Milk proteins in Food
2000
Shon Dong-Hwa | Kim Hyun-Jung Korea Food Research Institute, Songman (Korea Republic)
An enzyme-linked immunosorbent assay(ELISA) was developed for the detection of milk proteins in processed foods. The alpha s1-casein(alpha s1-CN), a heat stable major milk protein, was immunized into rabbits to produce specific antibodies. When competitive indirect ELISA(ciELISA) using anti-alpha s1-CN antibodies was established, its detection limit was 0.1 micro g/mL. The reactivities of the specific antibodies toward alpha s1-CN, skim milk, beta-CN and whey protein isolate(WPI) were 100, 37, 0.14 and 0.04%, respectively, as determined by ciELISA. However anti-alpha s1-CN antibodies did not have any reactivity to other milk proteins such as beta-lactoglobulin, alpha-lactalbumin, bovine serum albumin, and isolated soy protein. When sandwich ELISA was established, its detection limit was 0.01 micro g/mL which was 10 times more sensitive than that of ciELISA. In the spike test which was performed by adding 1-10% of whole CN to market milk, mean assay recovery as determined by sandwich ELISA was 94.8%(CV, 8.2%). Food stuffs and dairy products were assayed by sandwich ELISA to show 29, 0.13, 0.25, and 6.9% of whole CN in skim milk powder, WPI, semi-solid yoghurt, and processed cheese, respectively.
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