Production, characterization and use of five monoclonal antibodies to human IL-4.
We describe five murine monoclonal antibodies that are specific for human IL-4. At least three spatially distinct epitopes on the hIL-4 molecule are recognized by this panel of antibodies which allowed development of a sensitive sandwich EIA specific for hIL-4. The EIA is capable of detecting 200 pg/ml of hIL-4 and exhibits no detectable crossreactivity to seven other human cytokines examined. In addition to the EIA these antibodies are also useful in a number of other techniques for investigating hIL-4. Recombinant hIL-4 can be easily and efficiently purified by affinity chromatography using these mAbs. Western blotting with two of the antibodies detects as little as 1.8 ng/ml of hIL-4 in a 50 microliters sample. Finally, we present a method for cytoplasmic localization of hIL-4 by a fluorescence staining method utilizing a hIL-4 specific mAb.
Department of Pathology, Washington University School of Medicine, St. Louis, MO.
M G
MG
Scott
A D
AD
Levine
A W
AW
Butch
M B
MB
Bowen
K A
KA
Macke
M H
MH
Nahm
D000818:Animals; D000911:Antibodies, Monoclonal / Q000096:biosynthesis / Q000276:immunology*; D000918:Antibody Specificity; D015153:Blotting, Western; D002846:Chromatography, Affinity; D004797:Enzyme-Linked Immunosorbent Assay; D000939:Epitopes / Q000276:immunology; D006801:Humans; D007074:Immunoglobulin G / Q000276:immunology; D015847:Interleukin-4 / Q000032:analysis / Q000235:genetics / Q000276:immunology*; D007963:Leukocytes, Mononuclear / Q000378:metabolism; D008325:Mammary Neoplasms, Experimental; D051379:Mice; D008934:Mitogens / Q000494:pharmacology; D011994:Recombinant Proteins / Q000032:analysis / Q000276:immunology; D014162:Transfection; D014407:Tumor Cells, Cultured
D000911:Antibodies, Monoclonal; D000939:Epitopes; D007074:Immunoglobulin G; D008934:Mitogens; D011994:Recombinant Proteins; D015847:Interleukin-4