Mochizuki D Y, Eisenman J R, Conlon P J, Park L S, Urdal D L
J Immunol. 1986 May 15;136(10):3706-9.
The expression in yeast of a cDNA clone encoding murine granulocyte-macrophage colony-stimulating factor (GM-CSF) has made possible the purification of large quantities of this recombinant protein. Rabbits immunized with pure recombinant GM-CSF generated antibodies that were shown to be specific for both recombinant GM-CSF and GM-CSF isolated from natural sources. Other lymphokines, including IL 1 beta, IL 2, IL 3, and recombinant human GM-CSF did not react with the antiserum. The antiserum, together with recombinant GM-CSF that had been radiolabeled with 125I to high specific activity, formed the foundation for a rapid, sensitive, and quantitative radioimmunoassay specific for murine GM-CSF. Furthermore, the antiserum was found to inhibit the biologic activities of GM-CSF as measured in both a bone marrow proliferation assay and a colony assay, and thus should prove to be a useful reagent for dissecting the complex growth factor activities involved in murine hematopoiesis.
编码小鼠粒细胞-巨噬细胞集落刺激因子(GM-CSF)的cDNA克隆在酵母中的表达,使得大量纯化这种重组蛋白成为可能。用纯重组GM-CSF免疫的兔子产生了抗体,这些抗体被证明对重组GM-CSF和从天然来源分离的GM-CSF都具有特异性。其他细胞因子,包括IL-1β、IL-2、IL-3和重组人GM-CSF,均不与该抗血清发生反应。该抗血清与用125I标记至高比活性的重组GM-CSF一起,构成了一种针对小鼠GM-CSF的快速、灵敏且定量的放射免疫测定法的基础。此外,在骨髓增殖试验和集落试验中均发现该抗血清可抑制GM-CSF的生物学活性,因此应证明是剖析参与小鼠造血的复杂生长因子活性的有用试剂。