[Evaluation and determination of P-type-alpha amylase activity in human serum by monoclonal antibody]

We evaluated a kinetic procedure for determining pancreatic type of alpha-amylase isoenzyme (EC 3.2.1.1.) by using monoclonal salivary amylase antibody in human serum. The principle of this method is to measure residual pancreatic amylase activity after the reaction of monoclonal anti-salivary amylase antibody which is specifically inactivated human salivary amylase. The reaction of antibody with salivary alpha-amylase completed within 2 min of incubation and residual activity of pancreatic amylase was 100%, and the residual salivary amylase activity showed less than 3% of salivary amylase activity after incubation with antibody. The reproducibility and/or precision was 1.2% to 3.4% and the linearity obtained up to 1.500 U/l. Correlation coefficient between the proposed method (y) and electrophoresis (x1) and wheat germ inhibition method (x2) was well, respectively (r = 0.99 with both method, the regression curves was y = 1.02 x1-0.7 and y = 1.04x2-0.7). The reference value from healthy adults (male 98 and female 102) showed 13.6-48.2 U/l according to statistical parametric method and sex difference was not observed. The monoclonal anti-salivary amylase antibody was not cross-reacted with the other animal serum salivary type of amylase.