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Clinical Chemistry 35: 2169-2172, 1989;
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Clinical Chemistry, Vol 35, 2169-2172, Copyright © 1989 by American Association for Clinical Chemistry

Determination of alpha 2-macroglobulin-trypsin complex with a new synthetic substrate

K Kuroiwa, S Nakatsuyama, K Katayama and T Nagasawa
Nitto Boseki Co., Ltd., Biochemical Laboratory, Fukushima-Pref., Japan.

We have developed a colorimetric assay for quantifying alpha 2- macroglobulin-trypsin complex (alpha 2M-TRY) in human serum, based on use of a new chromogenic substrate D-gamma-tert-butyloxy-Glu-Gly-Arg-3- carboxy-4-hydroxyanilide dihydrochloride (PS-3001). Within-run CVs by this assay were 4.76%, 1.57%, and 0.83% for trypsin complex concentrations of 3.1, 12.2, and 48.1 U/L, respectively (n = 10 each). Between-day CVs were 5.38%, 3.12%, and 2.20% at each concentration, respectively (n = 7). Mean analytical recoveries of alpha 2M-TRY added to serum were 100%, 105%, and 101% for 9.2, 15.1, and 46.3 U/L, respectively (n = 2). The standard curve obtained was linear up to 330 U/L. We applied this method to the study of alpha 2M-TRY activity in sera from 97 healthy subjects (group 1), from 27 patients with acute pancreatitis (group 2), and from 25 patients with other chylopoietic diseases (group 3); results ranged from 0 to 1.2 U/L (mean = 0.5, SD = 0.3), from 1.2 to 77.4 U/L (mean = 14.6, SD = 19.0), and from 0 to 1.3 U/L (mean = 0.4, SD = 0.3), respectively. Concentrations of enzymatically active alpha 2M-TRY were significantly greater in sera from group 2 than in groups 1 and 3. The determination of serum alpha 2M-TRY activity by this simple, rapid, colorimetric method may be useful for the diagnosis and evaluation of pancreatic disease.




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Copyright © 1989 by the American Association for Clinical Chemistry.