Detection of human IL-1 alpha and IL-1 beta at the subpicomolar level by colorimetric sandwich enzyme immunoassay.
Two separate convenient sandwich enzyme immunoassay methods were developed for measuring the production of the monokines interleukin-1 alpha (IL-1 alpha) and interleukin-1 beta (IL-1 beta) from peripheral blood mononuclear cells. Polyclonal antisera raised against the recombinant proteins and selected on the basis of their ability to neutralize IL-1-induced IL-2 secretion were used for coating microtiter plates or preparing peroxidase-Fab' conjugates. Both techniques were able to accurately and specifically detect monokines from various sources in the sub-picomolar range and were not influenced by compounds currently used for cell activation. A high molecular weight form of IL-1 beta was demonstrated under certain conditions and the two enzyme immunoassays were successfully applied to the detection of IL-1 alpha and IL-1 beta present in cell supernatants following stimulation with mitogenic or chemical agents.
Ferrua
B
B
U 210 INSERM, Faculté de Médecine, Nice, France.
Becker
P
P
Schaffar
L
L
Shaw
A
A
Fehlmann
M
M
D000818:Animals; D000918:Antibody Specificity; D002474:Cell-Free System; D002850:Chromatography, Gel; D003124:Colorimetry / Q000592:standards; D003429:Cross Reactions; D003470:Culture Media; D000939:Epitopes / Q000032:analysis; D006801:Humans; D007106:Immune Sera; D007124:Immunoenzyme Techniques / Q000592:standards; D007375:Interleukin-1 / Q000032:analysis* / Q000276:immunology / Q000592:standards; D007963:Leukocytes, Mononuclear / Q000378:metabolism; D011817:Rabbits; D012015:Reference Standards; D012756:Sheep
D003470:Culture Media; D000939:Epitopes; D007106:Immune Sera; D007375:Interleukin-1