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抗白细胞介素1山羊抗体的制备:使用免疫吸附剂纯化白细胞介素1。

Preparation of goat antibodies against interleukin 1: use of an immunoadsorbent to purify interleukin 1.

作者信息

Mizel S B, Dukovich M, Rothstein J

出版信息

J Immunol. 1983 Oct;131(4):1834-7.

PMID:6352805
Abstract

Antibodies with specificity for interleukin 1 (IL 1) were produced in a goat immunized with purified IL 1 alpha obtained from the murine macrophage cell line, P388D1. The anti-IL 1 IgG were capable of completely inhibiting the biologic activity of IL 1 in the murine thymocyte assay but had no effect on IL 2-driven T cell responses. Although the anti-IL 1 IgG were produced by using mouse IL 1, these antibodies also recognized IL 1 prepared from a human monocyte leukemia cell line. An immunoadsorbent column was prepared and used for the large-scale purification of IL 1 in relatively high yield. Approximately 250 micrograms of purified IL 1 were obtained from 50 liters of culture fluid, the yield being 20% of the initial activity. Six major species of IL 1 were resolved by using tris-glycinate discontinuous polyacrylamide gels. The purified IL 1 exhibited 50% of its maximal activity in the thymocyte proliferation assay at a concentration of approximately 1 X 10(-11) M.

摘要

用从鼠巨噬细胞系P388D1获得的纯化白细胞介素1α(IL-1α)免疫山羊,产生了对白介素1(IL-1)具有特异性的抗体。抗IL-1 IgG在鼠胸腺细胞试验中能够完全抑制IL-1的生物活性,但对IL-2驱动的T细胞反应没有影响。尽管抗IL-1 IgG是用小鼠IL-1产生的,但这些抗体也能识别从人单核细胞白血病细胞系制备的IL-1。制备了免疫吸附柱并用于以相对高的产率大规模纯化IL-1。从50升培养液中获得了约250微克纯化的IL-1,产率为初始活性的20%。使用Tris-甘氨酸不连续聚丙烯酰胺凝胶分离出六种主要的IL-1。在胸腺细胞增殖试验中,纯化的IL-1在浓度约为1×10⁻¹¹ M时表现出其最大活性的50%。

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