Isolation and Measurement of Pancreatic Amylase in Human Serum and Urine

1972 ◽  
Vol 18 (12) ◽  
pp. 1493-1497 ◽  
Author(s):  
L Fridhandler ◽  
J Edward Berk ◽  
M Ueda
Keyword(s):  
Acute Pancreatitis ◽  
Human Serum ◽  
Fallopian Tube ◽  
Normal Subjects ◽  
Pancreatic Amylase ◽  
Salivary Amylase ◽  
Tissue Homogenates ◽  
Quantitative Procedure ◽  
Lines Of Evidence ◽  
Serum And Urine

Abstract We describe a sensitive quantitative procedure for separating isoamylases in human serum, urine, and tissue homogenates. Two components have been discerned with chromatographic characteristics resembling those of pancreatic and salivary amylases, respectively. Several lines of evidence—derived from studies in normal subjects, pancreatectomized patients, and patients with acute pancreatitis—indicate that the pancreas is probably the source of the component in serum and urine that exhibits characteristics of pancreatic amylase. The source of the component resembling salivary amylase has not yet been fully defined. Isoamylase analysis of extracts of fallopian tube and liver revealed two amylase components with chromatographic properties similar to pancreatic and salivary amylases, respectively.

scholarly journals Radioimmunoassay of Human Pancreatic Amylase with Monoclonal Antibody

1989 ◽  
Vol 26 (5) ◽  
pp. 416-421 ◽  
Author(s):  
Chiyo Fujita ◽  
Chihiro Kasai ◽  
Hiromi Kosuge ◽  
Kenji Ogata ◽  
Ichiyo Oshima ◽  
...  
Keyword(s):  
Monoclonal Antibody ◽  
Acute Pancreatitis ◽  
Normal Subjects ◽  
Enzymic Activity ◽  
Pancreatic Amylase ◽  
Normal Range ◽  
Salivary Amylase ◽  
Serum Samples ◽  
The Mean ◽  
Mean Concentration

A monoclonal antibody (E-21) was obtained that specifically binds to human pancreatic amylase and shows negligible cross-reaction with human salivary amylase. Using this antibody a radioimmunoassay was developed for pancreatic amylase in human serum. The assay was shown to be sensitive (detectable up to 7 mg/L), reproducible, and specific for pancreatic amylase. In normal subjects, the mean concentration of serum pancreatic amylase determined by this method was 36·3 mg/L with a 95% confidence range of 16·5 to 79·2 mg/L. A good correlation was observed between the concentrations of immunoreactive pancreatic amylase (IR-PA) and enzymatic activities in 20 serum samples ( r = 0·97). The concentration of serum IR-PA was below the detectable limit in pancreatectomised patients, and was greatly increased in patients with acute pancreatitis; the latter was accompanied by parallel changes in total enzymic activity. In patients with mumps, the serum IR-PA level was within the normal range whereas the total enzymic activity was elevated.

Specific immunoassay of alpha-amylase isoenzymes in human serum.

1985 ◽  
Vol 31 (8) ◽  
pp. 1331-1334 ◽  
Author(s):  
M Gerber ◽  
K Naujoks ◽  
H Lenz ◽  
W Gerhardt ◽  
K Wulff
Keyword(s):  
Monoclonal Antibody ◽  
Human Serum ◽  
Enzyme Immunoassay ◽  
Amylase Activity ◽  
Alpha Amylase ◽  
Pancreatic Amylase ◽  
Routine Method ◽  
Salivary Amylase ◽  
Amylase Isoenzymes

Abstract A monoclonal antibody (66C7) was prepared that specifically binds human salivary amylase (EC 3.2.1.1); it cross reacts with human pancreatic amylase by less than 1%. Two procedures are described for determination of isoamylases in human serum with this antibody: an enzyme immunoassay for determining amylase of salivary origin, and a routine method in which this amylase is immunoprecipitated and the remaining (pancreatic) amylase activity is assayed. Results by the two methods correlate well.

Automated measurement of amylase isoenzymes with 4-nitrophenyl-maltoheptaoside as substrate and use of a selective amylase inhibitor.

1984 ◽  
Vol 30 (7) ◽  
pp. 1219-1222 ◽  
Author(s):  
H Okabe ◽  
Y Uji ◽  
K Netsu ◽  
A Noma
Keyword(s):  
Standard Method ◽  
Selective Inhibitor ◽  
Pancreatic Amylase ◽  
Automated Measurement ◽  
Salivary Amylase ◽  
Amylase Inhibitor ◽  
Amylase Isoenzymes ◽  
Method R ◽  
Serum And Urine ◽  
Serum And Urine Samples

Abstract We automated a kinetic procedure for determining amylase isoenzymes in serum and urine samples. We used 4-nitro-phenylmaltoheptaoside as substrate and a selective amylase inhibitor with the Abbott-VP bichromatic system. By use of the maximum differences between pancreatic (P) and salivary (S) amylase activities remaining after inhibition by the selective inhibitor and by use of the linear range, a one-point standard method for calibration is proposed for determining amylase activities between about 50 and 1500 U/L when the P/S ratio exceeds 0.2. Results correlated well with those by electrophoresis and the Phadebas method (r = 0.99 for both pancreatic and salivary amylase). Reproducibilities (CVs) were 1.5% to 5.5% for pancreatic amylase and 1.4% to 3.3% for salivary amylase in serum, 0.8% to 2.0% for pancreatic amylase and 0.8% to 2.3% for salivary amylase in urine.

Immunocatalytic assay of pancreatic alpha-amylase in serum and urine with a specific monoclonal antibody.

1989 ◽  
Vol 35 (4) ◽  
pp. 662-664 ◽  
Author(s):  
E Svens ◽  
K Käpyaho ◽  
P Tanner ◽  
T H Weber
Keyword(s):  
Monoclonal Antibody ◽  
Reference Intervals ◽  
Cross Reactivity ◽  
Alpha Amylase ◽  
Healthy Controls ◽  
Pancreatic Amylase ◽  
Specific Monoclonal Antibody ◽  
Salivary Amylase ◽  
Patient Groups ◽  
Serum And Urine

Abstract In this immunocatalytic assay for alpha-amylase (EC 3.2.1.1) of pancreatic origin, a highly specific monoclonal antibody coupled to plastic beads is used to extract pancreatic amylase from samples, leaving salivary amylase in solution. The catalytic activity of the bound pancreatic amylase is then determined with blocked p-nitrophenyl maltoheptaoside as substrate. The method shows no cross-reactivity with salivary amylase, analytical recovery is 89-109% for pancreatic amylase, and interassay imprecision is 7.1-7.7%. We used the method to determine pancreatic amylase in serum and urine from healthy controls and different patient groups. The reference intervals for 34 supposedly healthy controls were: serum, 10-48 U/L (mean 27 U/L); urine, less than 20-435 U/L (mean 104 U/L). Results by the present assay correlated well with a salivary amylase inhibition assay (Boehringer Mannheim). We conclude that the described immunocatalytic assay is clinically useful for detecting increased activities of pancreatic amylase in serum and urine.

Fluorescence depolarization assay for quantifying alpha-amylase in serum and urine.

1985 ◽  
Vol 31 (9) ◽  
pp. 1478-1480 ◽  
Author(s):  
M Hofman ◽  
M Shaffar
Keyword(s):  
Human Serum ◽  
Calibration Curve ◽  
Fluorescence Polarization ◽  
Alpha Amylase ◽  
Pancreatic Amylase ◽  
Liquid Form ◽  
Fluorescence Depolarization ◽  
Endogenous Glucose ◽  
Hydrolysis Of ◽  
Serum And Urine

Abstract We have developed a new method for quantifying alpha-amylase (EC 3.2.1.1) in serum and urine by fluorescence depolarization. Amylase in the sample catalyzes the hydrolysis of the substrate, a fluorescein-labeled amylose. This results in decreased fluorescence polarization, owing to the increased rate of rotation of the amylose fragment relative to the intact substrate. The TDx amylase assay is calibrated with six human-serum-based pancreatic amylase calibrators. Amylase activities are determined by interpolation from the calibration curve, which is stored in the TDx analyzer's memory. Results correlate well with those by the Du Pont aca assay and the Beckman "DRI-STAT" assay. Endogenous glucose does not interfere. CVs are less than 6%, and the reagents are stable in liquid form.

Interaction of immobilized anti-salivary amylase antibody with human macroamylases: implications for use in a pancreatic amylase assay to distinguish macroamylasemia from acute pancreatitis.

1989 ◽  
Vol 35 (8) ◽  
pp. 1651-1654
Author(s):  
T E Mifflin ◽  
R W Forsman ◽  
D E Bruns
Keyword(s):  
Acute Pancreatitis ◽  
Healthy Volunteers ◽  
Amylase Activity ◽  
Electrophoretic Analysis ◽  
Pancreatic Amylase ◽  
Salivary Amylase ◽  
Isoenzyme Composition ◽  
The Mean

Abstract We examined the ability of an immobilized antibody to salivary amylase (Clin Chem 1985;33:1283-8) to react with amylase in macroamylasemic sera. The antibody removed 50% (SD 23%) of the total amylase activity from 39 macroamylase sera, a percentage indistinguishable (P greater than 0.75) from the percentage removed from concurrently analyzed sera from healthy volunteers (49%, SD 11%). Electrophoretic analysis of 23 macroamylasemic sera revealed that the antibody removed only part of the macroamylase band(s) in 71% of the cases. We conclude that the mean isoenzyme composition of the macroamylase complexes is essentially identical to the mean isoenzyme distribution in normal sera (i.e., about half salivary and half pancreatic amylase). Further, the immobilized antibody can be used to distinguish most patients with macroamylasemia from those with acute pancreatitis, because sera from the latter contain an increased proportion (greater than 80%) of pancreatic amylase.

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