Development and application of a sensitive radioimmunoassay for human granulocyte-macrophage colony-stimulating factor able to measure normal concentrations in blood.
A radioimmunoassay (RIA) for human granulocyte-macrophage colony-stimulating factor (GM-CSf) was developed based on antibodies from rabbits immunized with glycosylated recombinant human (rh) GM-CSF. The antibodies are specific for human GM-CSF and do not crossreact with other human hematopoietic growth factors or mouse GM-CSF. The antibodies also react with nonglycosylated rhGM-CSF, so E. coli-derived rhGM-CSF can be assayed as well. The RIA has a measuring range of about 10 to 200 pg/mL. Normal blood was found to contain 13 to 24 pg/mL (95% limits) with a mean of 18.5 pg/mL (n = 34). Monoclonal antibodies against GM-CSF could remove GM-CSF from normal human serum, thus ensuring that the GM-CSF measured in serum is real and does not represent nonspecific reactivity with our polyclonal rabbit antibodies. While previously published methods have been unable to measure GM-CSF in human serum under normal conditions, our more sensitive RIA does confirm the presence of small amounts of GM-CSF in serum or plasma and can therefore be used to detect fluctuations of GM-CSF in health and in disease.
Mortensen
B T
BT
Department of Internal Medicine/Hematology, Rigshospitalet, Copenhagen, Denmark.
Schifter
S
S
Pedersen
L B
LB
Jensen
A N
AN
Hovgaard
D
D
Nissen
N I
NI
D000818:Animals; D000906:Antibodies / Q000276:immunology; D000918:Antibody Specificity; D016466:CHO Cells; D006224:Cricetinae; D004355:Drug Stability; D005615:Freezing; D006031:Glycosylation; D016178:Granulocyte-Macrophage Colony-Stimulating Factor / Q000097:blood* / Q000276:immunology; D006358:Hot Temperature; D006801:Humans; D008832:Microchemistry; D011786:Quality Control; D011817:Rabbits / Q000276:immunology; D011863:Radioimmunoassay / Q000379:methods* / Q000592:standards; D011994:Recombinant Proteins / Q000276:immunology; D012016:Reference Values
D000906:Antibodies; D011994:Recombinant Proteins; D016178:Granulocyte-Macrophage Colony-Stimulating Factor