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    • 8. 发明授权
    • Enzymatic process for the determination of beta-lactam antibiotics
    • 用于测定β-内酰胺抗生素的酶法
    • US4546076A
    • 1985-10-08
    • US462233
    • 1983-01-31
    • Jacques DegelaenAlbert LoffetJean-Pierre Durieux
    • Jacques DegelaenAlbert LoffetJean-Pierre Durieux
    • G01N33/50C12N9/52C12N11/08C12Q1/04C12Q1/18C12Q1/34C12Q1/37C12R1/03C12Q1/36
    • C12Q1/34C12N11/08G01N2415/00
    • An enzymatic process for the determination of beta-lactam antibiotics in a biological liquid, comprising the steps of: (1) incubating the liquid with soluble D-alanyl-D-alanine-carboxypeptidase produced by Actinomadura R 39, said enzyme being immobilized on a water-insoluble support, the beta-lactam antibiotic reacting with the immobilized enzyme to form an inactive and equimolecular enzyme-antibiotic complex; (2) separating the immobilized enzyme from the liquid and washing it; (3) incubating the immobilized enzyme with a substrate solution to hydrolyze it and form an amount of D-alanine corresponding to the residual enzymatic activity; (4) determining the amount of D-alanine formed; and (5) comparing the determination of step (4) with a standard to obtain the concentration of the antibiotic in the biological liquid; as well as a test set for carrying out this process and comprising the necessary reagents.
    • 一种用于测定生物液体中β-内酰胺抗生素的酶促方法,包括以下步骤:(1)将该液体与由Actinomadura R 39产生的可溶性D-丙氨酰-D-丙氨酸 - 羧肽酶一起孵育,所述酶固定在 水不溶性载体,β-内酰胺抗生素与固定化酶反应形成无活性和等分子酶 - 抗生素复合物; (2)将固定化酶与液体分离并洗涤; (3)将固定化酶与底物溶液一起温育以使其水解并形成一定数量的对应于残留的酶活性的D-丙氨酸; (4)确定形成的D-丙氨酸的量; 和(5)将步骤(4)的测定与标准进行比较以获得生物液体中抗生素的浓度; 以及用于进行该方法并包含必要试剂的试验组。