使用高度吸附的抗干扰素，通过抗体亲和色谱法纯化人干扰素。

Purification of human interferon by antibody affinity chromatography, using highly absorbed anti-interferon.

作者信息

Berg K, Heron I, Hamilton R

出版信息

Scand J Immunol. 1978;8(5):429-36. doi: 10.1111/j.1365-3083.1978.tb00538.x.

DOI:10.1111/j.1365-3083.1978.tb00538.x

Abstract

Antibodies against partially purified human leucocyte interferon (PIF) were bound to Sepharose 4B and crude interferons applied on this affinity column were purified, up to 8 x 10(5) interferon units (IFU) per mg protein in one step. Antibodies against PIF were absorbed with immobilized crude human leucocyte interferon bound to Sepharose, whereby antibodies against impurities were predominantly removed. Extensively absorbed antisera were coupled to Sepharose and used for antibody affinity chromatography of crude interferon preparations. Leucocyte and fibroblast interferons were purified in one step with around 100% recovery, up to 1 x 10(8) IFU per mg protein, and Namalva interferon up to 2 x 10(7) IFU/mg. SDS electrophoresis of affinity-purified leucocyte interferon revealed that the interferon activity appeared in two bands (19,000 and 23,000 D).

摘要

将抗部分纯化人白细胞干扰素（PIF）的抗体与琼脂糖4B结合，将粗制干扰素应用于该亲和柱进行纯化，一步可纯化至每毫克蛋白质高达8×10⁵个干扰素单位（IFU）。抗PIF的抗体与结合在琼脂糖上的固定化粗制人白细胞干扰素结合而被吸收，从而主要去除了针对杂质的抗体。将大量吸收的抗血清与琼脂糖偶联，并用于粗制干扰素制剂的抗体亲和层析。白细胞干扰素和成纤维细胞干扰素一步纯化，回收率约为100%，每毫克蛋白质高达1×10⁸个IFU，而纳马夸干扰素高达2×10⁷个IFU/mg。亲和纯化的白细胞干扰素的SDS电泳显示，干扰素活性出现在两条带中（19,000和23,000 D）。