Rumschlag H S, Yakrus M A, Cohen M L, Glickman S E, Good R C
Division of Bacterial Diseases, Centers for Disease Control, Atlanta, Georgia 30333.
J Clin Microbiol. 1990 Mar;28(3):591-5. doi: 10.1128/jcm.28.3.591-595.1990.
A 35-kilodalton (kDa) recombinant antigen (35-kDa antigen) produced by Escherichia coli JM107 carrying DNA from Mycobacterium tuberculosis was purified and immunologically examined by in vivo and in vitro methods. A monoclonal antibody (2B2) was produced against the 35-kDa antigen. The protein was purified from the insoluble fraction of the recombinant E. coli strain by either affinity chromatography with the 2B2 monoclonal antibody or preparative isoelectric focusing. In enzyme-linked immunosorbent assay and Western blot (immunoblot) analyses, antibody to 2B2 reacted with whole-cell sonic extracts of M. tuberculosis and other slowly growing mycobacteria but not with two rapid growers, M. chelonae and M. fortuitum. An injection series totaling less than 1 mg of purified protein without adjuvant elicited a humoral response in guinea pigs. In one guinea pig, 10 micrograms of purified protein injected intradermally elicited both a humoral and a cell-mediated response. Results of these studies suggest that the 35-kDa antigen is a membrane-associated protein that stimulates both humoral and cell-mediated immune responses and should be evaluated as a vaccine candidate.
携带结核分枝杆菌DNA的大肠杆菌JM107产生的一种35千道尔顿（kDa）重组抗原（35-kDa抗原）被纯化，并通过体内和体外方法进行免疫学检测。产生了一种针对35-kDa抗原的单克隆抗体（2B2）。该蛋白通过用2B2单克隆抗体进行亲和层析或制备性等电聚焦从重组大肠杆菌菌株的不溶性部分中纯化出来。在酶联免疫吸附测定和蛋白质印迹（免疫印迹）分析中，2B2抗体与结核分枝杆菌和其他生长缓慢的分枝杆菌的全细胞超声提取物发生反应，但不与两种生长迅速的分枝杆菌，即龟分枝杆菌和偶然分枝杆菌发生反应。在豚鼠中，总量少于1毫克的纯化蛋白无佐剂注射系列引发了体液反应。在一只豚鼠中，皮内注射10微克纯化蛋白引发了体液和细胞介导的反应。这些研究结果表明，35-kDa抗原是一种与膜相关的蛋白，可刺激体液和细胞介导的免疫反应，应作为疫苗候选物进行评估。
