Application of a new monoclonal antibody for time-resolved fluoroimmunoassay of human pancreatic phospholipase A2.
A monoclonal antibody, designated 2E1, against human pancreatic phospholipase A2 was produced by hybridization of myeloma cells with spleen cells of immunized BALB/c mice. The hybridomas were screened for antibody production by time-resolved fluoroimmunoassay (TR-FIA). The antibody was found to belong to subclass I of murine IgG. The specificity of the antibody was confirmed by immunohistochemistry of pancreatic and other tissues, by immunoblotting of a crude aqueous extract of human pancreas and purified human pancreatic phospholipase A2 and by TR-FIA. A solid-phase time-resolved fluoroimmunoassay was developed by using the monoclonal anti-phospholipase A2 antibody as the catching antibody and a polyclonal sheep anti-phospholipase A2 antibody labelled with europium as the detecting antibody. The validity of the new TR-FIA of human pancreatic phospholipase A2 was confirmed by using it to measure the phospholipase A2 concentrations in serum samples from healthy subjects and from patients suffering from acute pancreatitis.
Santavuori
S A
SA
Labmaster Ltd., Turku, Finland.
Kortesuo
P T
PT
Eskola
J U
JU
Nevalainen
T J
TJ
D000208:Acute Disease; D000328:Adult; D000368:Aged; D000369:Aged, 80 and over; D000818:Animals; D000911:Antibodies, Monoclonal; D000918:Antibody Specificity / Q000276:immunology; D004591:Electrophoresis, Polyacrylamide Gel; D005260:Female; D015200:Fluoroimmunoassay / Q000379:methods*; D006801:Humans; D006825:Hybridomas / Q000276:immunology; D008297:Male; D051379:Mice; D008807:Mice, Inbred BALB C; D008875:Middle Aged; D010179:Pancreas / Q000201:enzymology*; D010195:Pancreatitis / Q000097:blood; D010741:Phospholipases A / Q000097:blood* / Q000276:immunology; D054467:Phospholipases A2; D015203:Reproducibility of Results; D012680:Sensitivity and Specificity
D000911:Antibodies, Monoclonal; D010741:Phospholipases A; D054467:Phospholipases A2