title: A cocktail of three monoclonal antibodies significantly increases the sensitivity of an enzyme immunoassay for human granulocyte-macrophage colony-stimulating factor.
abstract: A sensitive and specific two-site ELISA was developed for human granulocyte-macrophage colony-stimulating factor (huGM-CSF) based on monoclonal antibodies (mAbs) which have been selected for high affinities and different epitope specificities. Using a cocktail of three mAbs, both for coating and, in their labeled form, for detection, a major increase in sensitivity was achieved (20-fold) compared to a two-site assay employing two different mAbs (one for coating and one for detection). The assay is as sensitive as the most sensitive biological assays. Recombinant mammalian cell expressed and natural huGM-CSF can be reliably detected down to 100 pg/ml (7 pmol/l). In contrast to conventional bioassays, the ELISA is highly specific for huGM-CSF and does not detect other human lymphokines. Results from quantification of recombinant and natural huGM-CSF in ELISA and bioassay correlate.
authors.affiliation: Preclinical Research, Sandoz Pharma Ltd., Basle, Switzerland.
authors.forename: G
authors.initials: G
authors.lastname: Zenke
authors.forename: U
authors.initials: U
authors.lastname: Strittmatter
authors.forename: R
authors.initials: R
authors.lastname: Tees
authors.forename: E
authors.initials: E
authors.lastname: Andersen
authors.forename: B
authors.initials: B
authors.lastname: Fagg
authors.forename: H P
authors.initials: HP
authors.lastname: Kocher
authors.forename: M H
authors.initials: MH
authors.lastname: Schreier
mesh_terms: D000911:Antibodies, Monoclonal; D000915:Antibody Affinity; D000918:Antibody Specificity; D001681:Biological Assay; D004797:Enzyme-Linked Immunosorbent Assay / Q000379:methods*; D000939:Epitopes; D016178:Granulocyte-Macrophage Colony-Stimulating Factor / Q000032:analysis* / Q000276:immunology; D006801:Humans; D012680:Sensitivity and Specificity
chemical_list: D000911:Antibodies, Monoclonal; D000939:Epitopes; D016178:Granulocyte-Macrophage Colony-Stimulating Factor