Suzu S, Yanai N, Sato-Somoto Y, Yamada M, Kawashima T, Hanamura T, Nagata N, Takaku F, Motoyoshi K
Biochemical Research Laboratory, Morinaga Milk Industry Co, Ltd, Kanagawa, Japan.
Blood. 1991 May 15;77(10):2160-5.
We characterized the molecular species of human macrophage colony-stimulating factor (hM-CSF) found in serum and urine, using immunoblot analysis after partial purification on an antibody-bound affinity column. Although antibodies were prepared using the recombinant product of the large form of hM-CSF with a molecular weight (MW) of 85 Kd as the antigen, this immunoblot system was also capable of detecting the small form of hM-CSF with a MW of 40 to 60 Kd. A single band with a MW of 43 Kd, which reacted with anti-recombinant hM-CSF IgG but not with control IgG, was found when serum and urine from normal adults underwent electrophoresis on reduced sodium dodecyl sulfate-polyacrylamide gel and subsequent immunoblotting. This band represented a subunit of the large form of hM-CSF, because the large form of hM-CSF is a homodimer of a subunit with a MW of 43 Kd and the small form of hM-CSF is a homodimer of a subunit with a MW of 20 to 30 Kd. Analysis of serum and urine from leukemic patients and pregnant women, who had higher serum levels of hM-CSF than normal adults, showed only a single band with a MW of 43 Kd as a hM-CSF-specific molecule. These results suggest that the large form of hM-CSF is the major species in human body fluids.
我们通过在抗体结合亲和柱上进行部分纯化后使用免疫印迹分析，对血清和尿液中发现的人巨噬细胞集落刺激因子（hM-CSF）的分子种类进行了表征。尽管使用分子量（MW）为85 Kd的大形式hM-CSF的重组产物作为抗原制备了抗体，但该免疫印迹系统也能够检测到分子量为40至60 Kd的小形式hM-CSF。当正常成年人的血清和尿液在还原型十二烷基硫酸钠-聚丙烯酰胺凝胶上进行电泳并随后进行免疫印迹时，发现了一条分子量为43 Kd的单条带，它与抗重组hM-CSF IgG反应，但不与对照IgG反应。这条带代表大形式hM-CSF的一个亚基，因为大形式的hM-CSF是一个分子量为43 Kd的亚基的同二聚体，而小形式的hM-CSF是一个分子量为20至30 Kd的亚基的同二聚体。对白血病患者和孕妇的血清和尿液进行分析，这些患者的血清hM-CSF水平高于正常成年人，结果显示只有一条分子量为43 Kd的单条带作为hM-CSF特异性分子。这些结果表明，大形式的hM-CSF是人体体液中的主要种类。
