Radioimmunoassay for assessing exocrine pancreatic insufficiency, based on the differential enzymatic degradation of cobalamin-binding proteins.
Pancreatic proteases degrade the protein moiety of the R protein-cobalamin complex but not the intrinsic factor-cobalamin complex. Accordingly, we used these two proteins as substrates in an in vitro enzymatic assay to assess pancreatic function by incubating basal jejunal fluids with a mixture of intrinsic factor and cyano[57Co]cobalamin coupled to R-type protein and then using immunoprecipitation to determine the distribution of isotopically labeled cobalamin bound to the two proteins. With normal jejunal fluids, 91.2 (SD 6.1)% and 4.5 (SD 5.5)% of cyano[57Co]cobalamin was precipitated with antisera to intrinsic factor and anti-R protein, respectively. In the patients' jejunal fluids, the cyano[57Co]cobalamin precipitated with the respective antisera was 5.3 (SD 10.0)% and 96 (SD 6.2)%. In patients with other gastrointestinal problems, the sequestration of cobalamin was indistinguishable from that observed with the normal fluids. The clearcut discrimination this radioimmunoassay provided between abnormal and normal samples was confirmed by parallel comparative chromatographic analysis.
D002850:Chromatography, Gel; D010188:Exocrine Pancreatic Insufficiency / Q000175:diagnosis* / Q000378:metabolism; D006801:Humans; D007082:Ileum / Q000378:metabolism; D007408:Intestinal Absorption; D007419:Intestinal Secretions / Q000378:metabolism; D007437:Intrinsic Factor / Q000032:analysis / Q000378:metabolism; D007583:Jejunum / Q000378:metabolism; D010179:Pancreas / Q000201:enzymology*; D010447:Peptide Hydrolases / Q000378:metabolism*; D011863:Radioimmunoassay; D014155:Transcobalamins / Q000032:analysis / Q000378:metabolism*; D014805:Vitamin B 12 / Q000032:analysis / Q000378:metabolism
D014155:Transcobalamins; D007437:Intrinsic Factor; D010447:Peptide Hydrolases; D014805:Vitamin B 12