Ucb Societe Anonyme

Ucb Bioproducts, S.a.

Jacques Degelaen

Jean-Marie Frere

Albert Loffet

Jean-Pierre Durieux

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 produce by Actinomadura R39, the .beta.-lactam antibiotic reacting with the enzyme to form an inactive and equimolecular enzyme-antibiotic complex; (2) incubating the mixture obtained with a solution of a thioester substrate having the formula ##STR1## wherein R.sub.1 =benzoyl, phenylacetyl or N.sup..alpha. -acetyl-L-lysyl,

Enzymatic process for the determination of .beta.-lactam antibiotics

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.

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Genappe, Belgium

Jacques Degelaen