Development of a sandwich ELISA for ovine granulocyte/macrophage colony-stimulating factor.
Recombinant ovine granulocyte/macrophage colony-stimulating factor (rOv GM-CSF) has been expressed in Chinese hamster ovary cells. A stable, cloned line of these cells has been established which secretes high levels (40 mu g ml(-1)) of rOv GM-CSF. Three murine monoclonal antibodies (mAbs) were produced which reacted with rOv GM-CSF on Western blots. These mAbs also neutralised the activity of both recombinant and native Ov GM-CSF in a bone marrow haemopoietic progenitor cell assay. Two of the mAbs, which recognise mutually exclusive epitopes, were selected for the development of a sandwich enzyme-linked immunosorbent assay (ELISA) to measure GM-CSF in biological samples of ovine origin.
Entrican
G
G
Moredun Research Institute, Edinburgh, UK.
Deane
D
D
MacLean
M
M
Inglis
L
L
Thomson
J
J
McInnes
C
C
Haig
D M
DM
D000818:Animals; D000911:Antibodies, Monoclonal / Q000096:biosynthesis; D016466:CHO Cells; D006224:Cricetinae; D004797:Enzyme-Linked Immunosorbent Assay / Q000379:methods / Q000662:veterinary*; D015870:Gene Expression; D016178:Granulocyte-Macrophage Colony-Stimulating Factor / Q000032:analysis* / Q000235:genetics; D011994:Recombinant Proteins / Q000032:analysis / Q000235:genetics; D012756:Sheep / Q000378:metabolism*; D014162:Transfection
D000911:Antibodies, Monoclonal; D011994:Recombinant Proteins; D016178:Granulocyte-Macrophage Colony-Stimulating Factor