Detection of monocyte chemotactic and activating factor (MCAF) in normal blood and urine using a sensitive ELISA
| Autor: | Shingou Sakurai, Nobuo Ida, Genji Kawano |
|---|---|
| Rok vydání: | 1994 |
| Předmět: |
medicine.drug_class
medicine.medical_treatment Immunology Inflammation Enzyme-Linked Immunosorbent Assay Urine Biology Cross Reactions Monoclonal antibody Biochemistry Sensitivity and Specificity Mice Column chromatography Reference Values medicine Immunology and Allergy Animals Humans Molecular Biology Incubation Chemokine CCL2 Blood Specimen Collection Mice Inbred BALB C Chemotactic Factors Monocyte Antibodies Monoclonal Chemotaxis Hematology Molecular biology Cytokine medicine.anatomical_structure Cytokines Female Rabbits medicine.symptom |
| Zdroj: | Cytokine. 6(1) |
| ISSN: | 1043-4666 |
| Popis: | We developed a highly sensitive enzyme-linked immunosorbent assay (ELISA) for human monocyte chemotactic and activating factor (MCAF), an inflammatory cytokine that plays an important role in the recruitment of blood monocytes to areas of inflammation. The ELISA, which is based on a sandwich method using two newly-developed monoclonal antibodies, could quantitatively detect MCAF in the range between 2.5 pg/ml (50 fg/sample) to 300 pg/ml after incubation for a total of 2 h, and showed no cross-reactivity with various structurally-related IL-8 superfamily proteins. It was not affected by blood or urine components nonspecifically, and thus was directly applicable to clinical specimens. When serum and urine samples from healthy subjects were measured, they all turned out to contain detectable levels of MCAF (more than 30 pg/ml). By gel-filtration column chromatography analysis, MCAF in the body fluids was eluted as a single peak at the position corresponding to the molecular weight of 10 kD, suggesting that it exists as a monomer form, free from carrier proteins. The established ELISA here is expected to be effectively used for the further investigations on the relationship of MCAF with various inflammatory diseases. |
| Databáze: | OpenAIRE |
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