Bhunia A K, Johnson M G
Department of Food Science, University of Arkansas, Fayetteville 72703.
Appl Environ Microbiol. 1992 Jun;58(6):1924-9. doi: 10.1128/aem.58.6.1924-1929.1992.
A monoclonal antibody (MAb), EM-7G1, specific for Listeria monocytogenes was developed by using a previously developed MAb, C11E9 (A. K. Bhunia, P. H. Ball, A. T. Fuad, B. W. Kurz, J. W. Emerson, and M. G. Johnson, Infect. Immun. 59:3176-3184, 1991), to mask epitopes shared by L. monocytogenes and Listeria innocua in a 66-kDa cell surface protein. MAb EM-7G1 was an immunoglobulin subclass G1 antibody with kappa light chains. This MAb reacted with all 34 strains of L. monocytogenes tested and showed no cross-reaction with other Listeria spp. or other gram-positive or gram-negative organisms tested by enzyme-linked immunosorbent assay, dot blotting, and colony blotting. A second MAb, EM-6E11, reacted with all Listeria spp. tested but no other bacteria. In a Western blot (immunoblot) assay, EM-7G1 reacted with a crude cell surface protein of 66 kDa with a pI value of 6.7, while EM-6E11 reacted with two protein bands of 43 and 94 to 97 kDa with pI values of 4.0 and 4.3, respectively. Results with trypsin or pronase treatments indicated that the cell antigen reacting with EM-7G1 was on the surface of L. monocytogenes V7 and Scott A cells.
利用先前研制的单克隆抗体C11E9（A.K.布尼亚、P.H.鲍尔、A.T.富阿德、B.W.库尔兹、J.W.爱默生和M.G.约翰逊，《感染与免疫》59:3176 - 3184，1991年），开发出一种针对单核细胞增生李斯特菌的单克隆抗体（MAb）EM - 7G1，以掩盖单核细胞增生李斯特菌和无害李斯特菌在一种66 kDa细胞表面蛋白中共享的表位。MAb EM - 7G1是一种带有κ轻链的免疫球蛋白亚类G1抗体。该单克隆抗体与所测试的全部34株单核细胞增生李斯特菌发生反应，通过酶联免疫吸附测定、斑点印迹和菌落印迹法检测，未显示与其他李斯特菌属或其他革兰氏阳性或革兰氏阴性生物体发生交叉反应。第二种单克隆抗体EM - 6E11与所测试的所有李斯特菌属发生反应，但不与其他细菌发生反应。在蛋白质印迹（免疫印迹）分析中，EM - 7G1与一种66 kDa、pI值为6.7的粗制细胞表面蛋白发生反应，而EM - 6E11与两条分别为43 kDa和94至97 kDa、pI值分别为4.0和4.3的蛋白带发生反应。胰蛋白酶或链霉蛋白酶处理的结果表明，与EM - 7G1发生反应的细胞抗原位于单核细胞增生李斯特菌V7和斯科特A细胞的表面。
