Clinical Chemistry
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Clinical Chemistry 43: 569-574, 1997;
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(Clinical Chemistry. 1997;43:569-574.)
© 1997 American Association for Clinical Chemistry, Inc.

Articles

New sandwich ELISA for human urinary N-acetyl-ß-D-glucosaminidase isoenzyme B as a useful clinical test

Yoshito Numata1,a, Atsushi Morita1, Yoko Kosugi1, Kazunori Shibata1, Nozomu Takeuchi2 and Kiyohisa Uchida1

1 Diagnostic Science Department, Shionogi & Co., Ltd., 2-5-1 Mishima, Settsu-shi, Osaka 566, Japan.

2 Department of Clinical Laboratory Medicine, Ehime University School of Medicine, Shigenobu-cho, Ehime-Ken 791–02, Japan.
a Author for correspondence. Fax 06-319-4109.

We have developed a new ELISA for quantifying N-acetyl-ß-D-glucosaminidase (NAG) isoenzyme B in human urine after raising monoclonal antibodies against the isoenzyme from human placenta. Though the obtained antibodies reacted not only to isoenzyme B but also to A, we could detect isoenzyme B selectively by a two-step sandwich ELISA with a pair of selected antibodies at low pH in the first reaction. The detected limit was 0.5 µg/L for a sample volume of 25 µL. Within-run CVs ranged from 2.5% to 5.4% and between-run CVs ranged from 6.2% to 9.1%. Recoveries of NAG isoenzyme B added to each of three urine samples ranged from 91% to 114%. The dilution curves of urine samples showed good linearity. The cross-reactivity of NAG isoenzyme A was practically negligible (2–3%). The mean value for NAG isoenzyme B in spot urines from healthy adults was 2.9 µg/g creatinine. This ELISA method is rapid and precise enough for routine determination of NAG isoenzyme B in human urine.


Key Words: indexing terms: monoclonal antibody • renal damage • immunological activity




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