Production of genetically engineered biotinylated interleukin-2 and its application in a rapid nonradioactive assay for T-cell activation.
Production of genetically engineered biotinylated interleukin-2 and its application in a rapid nonradioactive assay for T-cell activation.
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基因工程生物素化白细胞介素 2 的生产及其在 T 细胞激活快速非放射性测定中的应用。
DOI：
10.1128/cdli.10.3.339-344.2003
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发表时间：
2003
期刊：
影响因子：
--
通讯作者：
Gosselin,EdmundJ
中科院分区：
文献类型：
--
作者：
Jordan,RobertA;Preissler,MarkT;Banas,JeffreyA;Gosselin,EdmundJ
The development of reliable assay systems that can measure lymphocyte activation in vitro has been a major goal of immunodiagnostics. Traditionally, tritiated thymidine incorporation has been used to monitor T-cell activation. Other methods include enzyme-linked immunosorbent assay (ELISA), enzyme-linked immunospot assay, and colorimetric assays. We have established a lymphocyte activation assay that utilizes fluorescein isothiocyanate (FITC)-streptavidin bound to recombinant biotinylated human interleukin-2 (IL-2). Utilizing recombinant DNA technology, a unique monobiotinylated human IL-2 has been created and isolated using the Promega PinPoint vector system. ELISA has been used to demonstrate streptavidin binding and recognition by a human IL-2-specific antibody. IL-2 function has been demonstrated using a murine IL-2-dependent T-cell line, CTLL-2, responsive to human IL-2. Recombinant biotinylated human IL-2 conjugated to streptavidin-FITC or streptavidin-horseradish peroxidase has been used to monitor T-cell activation in the presence of antigen as well as mitogen. The sensitivity and convenience of this method make this lymphocyte activation assay an attractive alternative to tritiated thymidine incorporation as a method for monitoring T-cell activation. In addition, the availability of a recombinant biotinylated human IL-2 will permit the production of a uniform product suitable for diagnostic and clinical application.
