Differentiation of human phospholipase A2 isoenzymes in serum and other body fluids with use of monoclonal antibodies.
Elevated phospholipase A2 activities in serum were measured in patients suffering from acute pancreatitis or various inflammatory diseases. The photometric phospholipase A assay of Hoffmann & Neumann (Klin. Wochenschr. 67 (1989) 106-109) was combined with immunoabsorption by different monoclonal antibodies directed against pancreatic phospholipase A2. Pancreatic phospholipase A2 was purified from human duodenal juice. Monoclonal antibodies were prepared by fusion of spleen cells from immunized mice with P3X63-Ag8-653 myeloma cells. Samples with phospholipase A2 activity were incubated in monoclonal antibody-coated microtitre plates. Phospholipase A2 activities were determined in the monoclonal antibody-treated samples as well as in control samples. The method allows the determination of the fraction of human phospholipase A2 isoenzymes in various biological materials. For pancreatic phospholipase A2 the specific binding capacity was about 60-80%, the unspecific binding was 5-30%. Practically no cross-reactivity was seen with partially purified serum phospholipase A2, with recombinant platelet phospholipase A2, or with the sera of patients with non-pancreatic diseases. In conclusion, the present study confirmed the presence of pancreatic phospholipase A2 in human duodenal juice and in the ascites of necrotizing pancreatitis. However, pancreatic isoenzyme was absent in non-pancreatic inflammatory diseases. Therefore, elevated phospholipase activities in non-pancreatic inflammatory diseases cannot be attributed to the pancreas.
Hiefinger-Schindlbeck
R T
RT
Institute for Clinical Chemistry, Bogenhausen Hospital, München, Germany.
Dasser
C
C
Hübner-Parajsz
C
C
Hoffmann
G E
GE
Guder
W G
WG
D000368:Aged; D000369:Aged, 80 and over; D000818:Animals; D000911:Antibodies, Monoclonal; D001665:Binding Sites; D001826:Body Fluids / Q000201:enzymology; D003429:Cross Reactions; D005260:Female; D006801:Humans; D007249:Inflammation / Q000201:enzymology*; D007527:Isoenzymes / Q000097:blood / Q000276:immunology / Q000302:isolation & purification / Q000378:metabolism*; D008297:Male; D051379:Mice; D008875:Middle Aged; D010179:Pancreas / Q000201:enzymology; D010195:Pancreatitis / Q000201:enzymology*; D010741:Phospholipases A / Q000097:blood / Q000276:immunology / Q000302:isolation & purification / Q000378:metabolism*; D054467:Phospholipases A2; D013154:Spleen / Q000166:cytology
D000911:Antibodies, Monoclonal; D007527:Isoenzymes; D010741:Phospholipases A; D054467:Phospholipases A2