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  • Title: High performance liquid chromatographic quantitation of carboxypeptidase activity secreted by human hep G2 cells.
    Author: Grimwood BG, Tarentino AL, Plummer TH.
    Journal: Anal Biochem; 1988 Apr; 170(1):264-8. PubMed ID: 2839052.
    Abstract:
    A new high performance liquid chromatographic method has been developed for the determination of carboxypeptidase N activity which quantitates the furylacryloyl-alanine released by enzymatic cleavage of furylacryloyl-alanyl-lysine or furylacryloyl-alanyl-arginine. A short isocratic gradient elutes the substrate and product in less than 7 min and multiple analyses are facilitated by an automatic sample injector. The microassay readily detects and quantitates carboxypeptidase N activity secreted into culture medium. It was determined that approximately 1 X 10(6) Hep G2 cells at early confluence secreted 1 ng of carboxypeptidase N in 24 h. The microassay will also detect as little as 51 pg of purified carboxypeptidase N or 8 pg of carboxypeptidase B.
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