A sensitive, rapid assay for the detection of human granulocyte-macrophage colony stimulating factor.
Several monoclonal antibodies were developed against recombinant human granulocyte colony stimulating factor (hu-GM-CSF). All were reactive to the protein by enzyme linked immunoabsorbent assay (ELISA), and one, 1G2 was capable of immunoprecipitating significant levels of radiolabeled hu-GM-CSF. When 1G2 and a second GM-CSF reactive monoclonal antibody, 3G11, were used in a double determinant assay, the level of detection of hu-GM-CSF in solution was approximately 500 ng/ml. Additional sensitivity was gained by using affinity purified rabbit polyclonal antibodies, together with the monoclonal 1G2. Using such a configuration in a double determinant assay one could detect 10-90 ng/ml of human-GM-CSF in solution with no reactivity observed to CSF-1 or granulocyte-CSF.
Conlon
P J
PJ
Immunex Corporation, Seattle, WA 98101.
McMasters
D
D
Prickett
K S
KS
D000911:Antibodies, Monoclonal; D011232:Chemical Precipitation; D003115:Colony-Stimulating Factors / Q000032:analysis* / Q000276:immunology; D004797:Enzyme-Linked Immunosorbent Assay; D016178:Granulocyte-Macrophage Colony-Stimulating Factor; D006133:Growth Substances / Q000032:analysis* / Q000276:immunology; D006801:Humans; D007118:Immunoassay / Q000379:methods*; D007074:Immunoglobulin G
D000911:Antibodies, Monoclonal; D003115:Colony-Stimulating Factors; D006133:Growth Substances; D007074:Immunoglobulin G; D016178:Granulocyte-Macrophage Colony-Stimulating Factor