Purification of hemopoietin 1: a multilineage hemopoietic growth factor.

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Abstract
Hemopoietin 1 (H-1) and the mononuclear phagocyte specific growth factor CSF-1 [colony-stimulating factor] act synergistically on developmentally early bone marrow cells to generate primitive CSF-1 receptor-bearing cells. The H-1 activity of the serum-free medium conditioned by the human urinary bladder carcinoma cell line 5637 was shown to result from the sum of the activities of 2 charged species (pI .apprxeq. 4.8, .apprxeq. 85%; pI .apprxeq. 5.3, .apprxeq. 15%) of similar size. No qualitative difference in the biological activity of these 2 species was detected. A 4-step procedure, involving batch DEAE-cellulose chromatography, chromatofocusing, gel filtration and hydrophobic chromatography was developed for the major (pI .apprxeq. 4.8) species. H-1 was purified .apprx. 65,000-fold and recovered as 32% of the total activity of the starting material. The lowest concentration yielding maximal biological activity was .apprxeq. 0.25 ng/ml. The 125I-labeled purified H-1, in either native or reduced form, behaved as a homogeneous single band that coelectrophoresed with the biological activity of purified H-1 on sodium dodecyl sulfate gel electrophoresis (NaDodSO4/PAGE). The molecular mass of the purified reduced H-1, determined by NaDodSO4/PAGE was .apprxeq. 17kDa [kilodalton]. Evidently, the purified H-1 is a multilineage hemopoietic growth factor.

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