期刊名称：Proceedings of the National Academy of Sciences
印刷版ISSN：0027-8424
电子版ISSN：1091-6490
出版年度：1982
卷号：79
期号：2
页码：601-605
DOI：10.1073/pnas.79.2.601
语种：English
出版社：The National Academy of Sciences of the United States of America
摘要：Leukocyte inhibitory factor (LIF), a lymphokine that inhibits the random and directed migration of polymorphonuclear (PMN) leukocytes, was purified from a human non-T, non-B leukemia cell line (Reh). From 10 liters of serum-free supernatant, 1.3 microgram of protein with LIF activity was obtained by the sequential use of affinity chromatography with concanavalin A-Sepharose, hydrophobic chromatography with hexylagarose, and gel filtration chromatography. The specific activity of LIF recovered represented an 80,000-fold purification over that of the initial crude serum-free supernatants, and the preparation at that point was estimated to be 80--90% pure. To both assess the purity of the preparation and provide a further purification step, Reh LIF activity recovered by the above procedures was subjected to isoelectric focusing. One major stainable protein band was identified; its isoelectric point was pH 5.4--5.5. Gels run in parallel for recovery of biologic activity revealed only one region (pH 5.4--5.5) with ability to inhibit PMN leukocyte migration. Iodination of Reh LIF resulted in a loss of biologic activity, but isoelectric focusing of this material revealed one major 125I-labeled band (pH 5.1) and several minor bands. The coincidence of biologic LIF activity with one stainable protein band as identified by isoelectric focusing implies that the final product may be homogeneous.
