title: Detection of human elastase isoforms by the ScheBo Pancreatic Elastase 1 Test.
abstract: Determination of fecal pancreatic elastase content by ELISA is a reliable, noninvasive clinical test for assessing exocrine pancreatic function. Despite the widespread use of commercial tests, their exact molecular targets remain poorly characterized. This study was undertaken to clarify which human pancreatic elastase isoforms are detected by the ScheBo Pancreatic Elastase 1 Stool Test and whether naturally occurring genetic variants influence the performance of this test. Using recombinantly expressed and purified human pancreatic proteinases, we found that the test specifically measured chymotrypsin-like elastases (CELA) 3A and 3B (CELA3A and CELA3B), while CELA2A was not detected. Inactive proelastases, active elastases, and autolyzed forms were detected with identical efficiency. CELA3B elicited approximately four times higher ELISA signal than CELA3A, and we identified Glu154in CELA3B as the critical determinant of detection. Common genetic variants of CELA3A and CELA3B had no effect on test performance, with the exception of the CELA3B variant W79R, which increased detection by 1.4-fold. Finally, none of the human trypsin and chymotrypsin isoforms were detected. We conclude that the ScheBo Pancreatic Elastase 1 Stool Test is specific for human CELA3A and CELA3B, with most of the ELISA signal attributable to CELA3B.NEW & NOTEWORTHYThe ScheBo Pancreatic Elastase 1 Stool Test is widely used to assess pancreatic exocrine function, yet its molecular targets have been poorly defined. We demonstrate that, among the human pancreatic proteinases, the test measures the elastase isoform CELA3B and, to a lesser extent, CELA3A. Genetic variants of the human CELA3 isoforms have no significant effect on test performance.
authors.affiliation: Center for Exocrine Disorders, Department of Molecular and Cell Biology, Boston University Henry M. Goldman School of Dental Medicine, Boston, Massachusetts.|Hungarian Academy of Sciences Momentum Gastroenterology Multidisciplinary Research Group, University of Szeged, Szeged, Hungary; and.
authors.forename: Anna Zsófia
authors.initials: AZ
authors.lastname: Tóth
authors.affiliation: Center for Exocrine Disorders, Department of Molecular and Cell Biology, Boston University Henry M. Goldman School of Dental Medicine, Boston, Massachusetts.
authors.forename: András
authors.initials: A
authors.lastname: Szabó
authors.affiliation: Center for Exocrine Disorders, Department of Molecular and Cell Biology, Boston University Henry M. Goldman School of Dental Medicine, Boston, Massachusetts.
authors.forename: Eszter
authors.initials: E
authors.lastname: Hegyi
authors.affiliation: Hungarian Academy of Sciences Momentum Gastroenterology Multidisciplinary Research Group, University of Szeged, Szeged, Hungary; and.|Institute for Translational Medicine and First Department of Medicine, University of Pécs, Pécs, Hungary.
authors.forename: Péter
authors.initials: P
authors.lastname: Hegyi
authors.affiliation: Center for Exocrine Disorders, Department of Molecular and Cell Biology, Boston University Henry M. Goldman School of Dental Medicine, Boston, Massachusetts; miklos@bu.edu.
authors.forename: Miklós
authors.initials: M
authors.lastname: Sahin-Tóth
mesh_terms: D004796:Clinical Enzyme Tests / Q000379:methods*; D004797:Enzyme-Linked Immunosorbent Assay; D005243:Feces / Q000201:enzymology; D014644:Genetic Variation; D057809:HEK293 Cells; D006801:Humans; D007527:Isoenzymes; D010196:Pancreatic Elastase / Q000032:analysis* / Q000235:genetics; D011237:Predictive Value of Tests; D011933:Reagent Kits, Diagnostic; D015203:Reproducibility of Results; D014162:Transfection
chemical_list: D007527:Isoenzymes; D011933:Reagent Kits, Diagnostic; C000617121:CELA3A protein, human; C000617122:CELA3B protein, human; D010196:Pancreatic Elastase
keywords: chronic pancreatitis; digestive proteinase; elastase; pancreas; pancreatic insufficiency