Omori F, Okamura S, Hayashi S, Yamaga S, Hirota Y, Niho Y
Cancer Center, Faculty of Medicine, Kyushu University, Fukuoka, Japan.
Biotherapy. 1989;1(3):161-7. doi: 10.1007/BF02170885.
An IgG monoclonal antibody against recombinant human granulocyte-macrophage colony-stimulating factor (GM-CSF), designated HGM1, was produced by fusion of immune mouse splenocytes with HAT-sensitive murine myeloma cells. A sandwich enzyme-linked immunosorbent assay (ELISA) for measurement of human GM-CSF was developed using this HGM1 and a polyclonal antibody against GM-CSF raised in a rabbit. GM-CSF in culture supernatants of phytohemagglutinin (PHA)- or concanavalin A (Con A)-stimulated peripheral blood mononuclear cells (PBMC) were measured by this ELISA system and the conventional CFU-GM colony formation method. The data indicated that the ELISA was highly efficient and sensitive for the detection of as little as 50 pg/ml recombinant GM-CSF. The CFU-GM colony assay may be influenced by other cytokines which can enhance or suppress colony formation, and ELISA for GM-CSF is more useful for kinetic studies of precise levels of production from PBMC.
通过将免疫小鼠脾细胞与对氨基蝶呤（HAT）敏感的鼠骨髓瘤细胞融合，制备了一种针对重组人粒细胞巨噬细胞集落刺激因子（GM-CSF）的IgG单克隆抗体，命名为HGM1。使用该HGM1和在兔体内产生的抗GM-CSF多克隆抗体，开发了一种用于检测人GM-CSF的夹心酶联免疫吸附测定（ELISA）。用该ELISA系统和传统的CFU-GM集落形成方法，测定了植物血凝素（PHA）或刀豆蛋白A（Con A）刺激的外周血单个核细胞（PBMC）培养上清液中的GM-CSF。数据表明，该ELISA对低至50 pg/ml的重组GM-CSF的检测具有高效性和敏感性。CFU-GM集落测定可能会受到其他可增强或抑制集落形成的细胞因子的影响，而GM-CSF的ELISA对于精确测定PBMC产生水平的动力学研究更有用。
