Evanoff H L, Burdick M D, Moore S A, Kunkel S L, Strieter R M
Department of Pathology, University of Michigan Medical School, Ann Arbor 48109-0360.
Immunol Invest. 1992 Feb;21(1):39-45. doi: 10.3109/08820139209069361.
The recruitment of monocytes into tissue is associated with both acute and chronic inflammation. Although monocyte migration is measured in vitro by monocyte chemotaxis, this technique is often difficult to determine the specific quantitative contribution of a monocyte chemotaxin. We have developed a sensitive sandwich ELISA for the detection of monocyte chemoattractant protein-1 (MCP-1), a highly specific monocyte activating/chemotactic peptide. Polyclonal antibodies were generated from rabbits. The IgG fraction of the antiserum was isolated by a protein A column, with a portion of the antibodies biotinylated. Avidin-conjugated horse radish peroxidase was used for enzymatic, colorimetric analysis. The lower threshold for detection of MCP-1 was 50 pg/ml, and the ELISA was specific for MCP-1, since it failed to recognize other cytokines in a dose-dependent fashion. Furthermore, this ELISA had the capacity to measure endothelial cell and pulmonary fibroblast-derived MCP-1. The development of a sensitive ELISA for the detection MCP-1 is significant, since it will allow the measurement MCP-1 from biologically relevant fluids, and aid in establishing whether MCP-1 is present in disease.
单核细胞募集到组织中与急性和慢性炎症均相关。尽管单核细胞迁移在体外通过单核细胞趋化作用来测定，但该技术往往难以确定单核细胞趋化因子的具体定量贡献。我们已经开发出一种灵敏的夹心酶联免疫吸附测定法（ELISA），用于检测单核细胞趋化蛋白-1（MCP-1），这是一种高度特异性的单核细胞激活/趋化肽。用兔子制备多克隆抗体。抗血清的IgG部分通过蛋白A柱分离，一部分抗体用生物素标记。抗生物素蛋白偶联的辣根过氧化物酶用于酶促比色分析。检测MCP-1的下限为50 pg/ml，且该ELISA对MCP-1具有特异性，因为它不能以剂量依赖方式识别其他细胞因子。此外，该ELISA能够检测内皮细胞和肺成纤维细胞来源的MCP-1。开发一种用于检测MCP-1的灵敏ELISA具有重要意义，因为它将能够从生物学相关液体中检测MCP-1，并有助于确定疾病中是否存在MCP-1。
