Production of polyclonal and monoclonal antibodies to human granulocyte colony-stimulating factor (GCSF) and development of immunoassays.
Murine monoclonal antibodies and a sheep polyclonal antiserum against recombinant human granulocyte colony-stimulating factor (GCSF) have been produced. These have been used to develop an immunoassay which can detect 250 pg/ml (25 U) of both natural and recombinant human GCSF. The assay involves forming a complex between GCSF and a monoclonal anti-GCSF, binding of the complex to microtitre wells coated with sheep anti-GCSF and detection of the bound complex with 125I-labelled sheep anti-mouse IgG. Unlike the classical bone marrow assay and other cell line based bioassays for GCSF, the immunoassay was specific for the cytokine, showing no cross-reactivity with GM-CSF, IL-6, IL-3 or IL-1 alpha and -beta. The assay does not exhibit interfering matrix effects when used for the estimation of human GCSF in serum.
National Institute for Biological Standards and Control, Herts, U.K.
M
M
Wadhwa
R
R
Thorpe
C R
CR
Bird
A J
AJ
Gearing
D000818:Animals; D000906:Antibodies / Q000276:immunology / Q000302:isolation & purification*; D000911:Antibodies, Monoclonal / Q000276:immunology / Q000302:isolation & purification*; D002460:Cell Line; D003115:Colony-Stimulating Factors / Q000032:analysis* / Q000276:immunology; D016179:Granulocyte Colony-Stimulating Factor; D006801:Humans; D015151:Immunoblotting; D051379:Mice; D008807:Mice, Inbred BALB C; D011863:Radioimmunoassay / Q000379:methods*; D012756:Sheep
D000906:Antibodies; D000911:Antibodies, Monoclonal; D003115:Colony-Stimulating Factors; D016179:Granulocyte Colony-Stimulating Factor