一种抗人白细胞干扰素单克隆抗体在人α干扰素纯化及放射免疫测定中的应用。

Use of an anti-human leukocyte interferon monoclonal antibody for the purification and radioimmunoassay of human alpha interferon.

作者信息

Meurs E, Rougeot C, Svab J, Laurent A G, Hovanessian A G, Robert N, Gruest J, Montagnier L, Dray F

出版信息

Infect Immun. 1982 Sep;37(3):919-26. doi: 10.1128/iai.37.3.919-926.1982.

DOI:10.1128/iai.37.3.919-926.1982

PMID:6182105

原文链接:https://pmc.ncbi.nlm.nih.gov/articles/PMC347626/

Abstract

Mouse monoclonal antibody directed against human leukocyte alpha interferon (IFN-alpha) was coupled to Sepharose and used as an immunoadsorbent to purify human IFN-alpha. Leukocyte and lymphoblastoid (Namalva) IFNs were retained by the immunoadsorbent with a specificity of 80 to 100% and 40 to 60%, respectively. Human IFN-beta or -gamma and mouse IFN were not retained. The purified IFN-alpha retained its antiviral and anticellular properties as well as its ability to induce the 2-5A synthetase in human cells with a specific activity similar to that of the crude IFN. Sodium dodecyl sulfate-polyacrylamide gel electrophoresis of radioactively labeled IFN showed that it consisted of several proteins in the molecular weight range of 17,000 to 27,000. 125I-labeled IFN-alpha with a high specific activity (2,000 Ci/mmol) was used in a radioimmunoassay for the titration of IFN-alpha.

摘要

针对人白细胞α干扰素（IFN-α）的小鼠单克隆抗体与琼脂糖凝胶偶联，并用作免疫吸附剂来纯化人IFN-α。白细胞干扰素和淋巴母细胞样（Namalva）干扰素分别以80%至100%和40%至60%的特异性被免疫吸附剂保留。人IFN-β或-γ以及小鼠干扰素未被保留。纯化后的IFN-α保留了其抗病毒和抗细胞特性，以及在人细胞中诱导2-5A合成酶的能力，其比活性与粗制干扰素相似。放射性标记的IFN的十二烷基硫酸钠-聚丙烯酰胺凝胶电泳表明，它由几种分子量在17,000至27,000范围内的蛋白质组成。具有高比活性（2,000 Ci/mmol）的125I标记的IFN-α用于放射免疫测定以滴定IFN-α。