Development of an indirect method for measuring porcine pancreatic lipase in human duodenal fluid.
Patients with exocrine pancreatic insufficiency are usually treated with porcine pancreatic enzymes but the bioavailability of these enzymes in the gut remains a matter of discussion. In order to determine the duodenal availability of porcine pancreatic lipase (PPL) present in pancreatic extracts (PE) taken orally, we developed a method for quantifying PPL in samples containing both PPL and human pancreatic lipase (HPL). Total pancreatic lipase activity measurements using the pH-stat technique and tributyrin as substrate were combined with an HPL-specific ELISA. Based on the known specific activity of the purified HPL, its activity was deduced from the ELISA measurements, and the PPL activity was obtained by subtracting the HPL activity from the total pancreatic lipase activity. This assay was established and validated using various samples containing pure PPL and recombinant HPL or PE, mixed or not with human duodenal juice. Samples collected in vivo from patients treated with PE were also tested. It was found that PPL did not affect the HPL ELISA, and the indirect PPL assay gave a measurement accuracy of 6.6% with the samples containing pure PPL and 10% with those containing PE. This assay was also used successfully to discriminate between PPL and the endogenous HPL present in the duodenal contents of patients with severe pancreatic insufficiency treated with PE. This method might provide a useful means of assessing the availability of PEs at their site of action, in the absence of a PPL-specific ELISA.
Tuvignon
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N
Laboratoire d'Enzymologie Interfaciale et de Physiologie de la Lipolyse, CNRS UPR 9025, IBSM, 31 Chemin Joseph-Aiguier, 13009 Marseille Cedex 20, France.
Abousalham
A
A
Tocques
F
F
De Caro
J
J
De Caro
A
A
Laugier
R
R
Carrière
F
F
D000818:Animals; D000911:Antibodies, Monoclonal / Q000276:immunology; D000918:Antibody Specificity; D001826:Body Fluids / Q000737:chemistry*; D004063:Digestion; D004386:Duodenum / Q000378:metabolism*; D004797:Enzyme-Linked Immunosorbent Assay; D005502:Food; D006801:Humans; D008049:Lipase / Q000032:analysis* / Q000276:immunology / Q000378:metabolism; D010179:Pancreas / Q000201:enzymology*; D011994:Recombinant Proteins / Q000032:analysis / Q000276:immunology / Q000378:metabolism; D015203:Reproducibility of Results; D012680:Sensitivity and Specificity; D045602:Steatorrhea / Q000628:therapy; D013552:Swine / Q000276:immunology
D000911:Antibodies, Monoclonal; D011994:Recombinant Proteins; D008049:Lipase