Rodríguez E, Martín R, García T, Hernández P E, Sanz B
Departamento de Higiene y Tecnología de los Alimentos, Facultad de Veterinaria, Universidad Complutense, Madrid, Spain.
J Dairy Res. 1990 May;57(2):197-205. doi: 10.1017/s0022029900026807.
An indirect enzyme-linked immunosorbent assay was successfully developed for the detection of defined amounts of cows' milk (1-50%) in sheeps' milk and cheese. The assay used polyclonal antibodies raised in rabbits against bovine caseins (BC). The anti-BC antibodies were recovered from the crude antiserum by immunoadsorption and elution from a column containing BC. The antibodies were biotinylated and rendered cows' milk specific by mixing them with lyophilized ovine and caprine caseins. ExtrAvidin-peroxidase was used to detect the biotinylated anti-BC antibodies bound to BC immobilized on 96-well plates. Subsequent enzymic conversion of substrate gave clear absorbance differences when assaying mixtures of sheeps' milk and cheese containing variable amounts of cows' milk.
成功开发了一种间接酶联免疫吸附测定法，用于检测羊奶和奶酪中特定含量（1%-50%）的牛奶。该测定法使用兔抗牛酪蛋白（BC）产生的多克隆抗体。通过免疫吸附从粗抗血清中回收抗BC抗体，并从含有BC的柱上洗脱。将抗体生物素化，并通过与冻干的羊和山羊酪蛋白混合使其对牛奶具有特异性。使用抗生物素蛋白-过氧化物酶检测与固定在96孔板上的BC结合的生物素化抗BC抗体。在检测含有不同含量牛奶的羊奶和奶酪混合物时，随后底物的酶促转化产生了明显的吸光度差异。
