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Fluorometric microassay of trypsin and enteropeptidase in children — comparison with a titrimetic assay
Author(s) -
MITCHELL JOHN D.,
MESSINA ISABELLA M.
Publication year - 1984
Publication title -
journal of paediatrics and child health
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.631
H-Index - 76
eISSN - 1440-1754
pISSN - 1034-4810
DOI - 10.1111/j.1440-1754.1984.tb00101.x
Subject(s) - trypsin , chromatography , medicine , enteropeptidase , hydrochloride , substrate (aquarium) , enzyme , microbiology and biotechnology , biochemistry , chemistry , biology , recombinant dna , ecology , fusion protein , gene
Two trypsin assay methods for the estimation of this enzyme in duodenal fluid from children have been compared. Assay results for a fluorometric method based on the use of N‐carbobenzoxy‐diglycyl‐L‐arginyl‐2‐naphthylamide hydrochloride (GANA) as the trypsin substrate were found to correlate well (r=0.91, P<0.001) with those obtained with a much less sensitive titrimetric assay which used benzoylarginine ethylester hydrochloride (BAEE) as substrate. The higher sensitivity of the fluorometric assay has allowed accurate determination of trypsin activity in 10 μl aliquots of duodenal fluid. This low volume requirement makes the assay suitable for studies on infants of all ages and conserves duodenal fluid for use in other investigations often warranted during the assessment of childhood malabsorption. The fluorometric assay has also been used to monitor the separation of enteropeptidase from trypsin(ogen) by chromatography on Sephacryl S‐200 in samples of duodenal fluid from two children. Different proteolytic pathway deficiencies were confirmed in these children.