Villarreal-Ramos B, Sanchez-Garcia J, Stoker N, Timms E, Chomer D, Raff K, Mitchison N A
Imperial Cancer Research Fund, Department of Biology, Medawar University College, London, GB.
Eur J Immunol. 1991 Oct;21(10):2621-4. doi: 10.1002/eji.1830211047.
Parasite expression libraries have so far been screened with antibodies, DNA probes or T cell clones. Immunity to many parasites, such as Mycobacterium leprae, is largely mediated by T cells, and so the screening of such libraries for T cell epitopes is an important step toward the development of effective vaccines and diagnostic reagents. A new method for screening of lambda gt11 libraries with uncloned T cell populations is presented here, which takes advantage of the fact that the recombinant proteins contain beta-galactosidase as their leader peptide; this allows them to be semipurified by means of anti-beta-galactosidase antibodies coated on the bottom of microtiter plate wells, within which a proliferation assay can then be carried out. Optimum conditions for the assay were determined, using the M. leprae 18-kDa antigen as a test antigen.
迄今为止,已利用抗体、DNA探针或T细胞克隆对寄生虫表达文库进行筛选。对许多寄生虫(如麻风分枝杆菌)的免疫很大程度上由T细胞介导,因此筛选此类文库以寻找T细胞表位是开发有效疫苗和诊断试剂的重要一步。本文介绍了一种用未克隆的T细胞群体筛选λgt11文库的新方法,该方法利用了重组蛋白含有β-半乳糖苷酶作为其前导肽这一事实;这使得它们能够通过包被在微量滴定板孔底部的抗β-半乳糖苷酶抗体进行半纯化,然后在其中进行增殖测定。以麻风分枝杆菌18 kDa抗原作为测试抗原,确定了该测定的最佳条件。