Monoclonal Antibodies Against a Mycobacterium tuberculosis Ag85B-Hsp16.3 Fusion Protein
Monoclonal Antibodies Against a Mycobacterium tuberculosis Ag85B-Hsp16.3 Fusion Protein
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抗结核分枝杆菌 Ag85B-Hsp16.3 融合蛋白的单克隆抗体
DOI：
10.1089/hyb.2011.0047
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发表时间：
2011-10-01
期刊：
影响因子：
--
通讯作者：
Xu, Zhikai
中科院分区：
文献类型：
--
作者：
Zhao, Shanmin;Shi, Jieran;Xu, Zhikai
结核分枝杆菌分泌型蛋白Ag85B和Hsp16.3是近年来研究的热点。这些蛋白对细菌阴性结核病(TB)患者具有很高的敏感性，对细菌阴性结核病的快速诊断有价值。融合蛋白包括多个抗原，如Ag85B和Hsp16.3，与单一抗原相比，可提高诊断活动性结核病的敏感性和特异性。许多研究表明，识别特定的结核分枝杆菌抗原的单抗的产生和基于单抗的检测在结核病的阳性检测中被发现是有价值的，特别是对涂阳肺结核。一些单抗目前被用于结核病的血清诊断。因此，本研究利用大肠杆菌系统对Ag85B-Hsp16.3融合蛋白进行了表达和纯化。用常规的小鼠杂交瘤技术制备了3株Ag85B-Hsp16.3融合蛋白特异性单抗。用酶联免疫吸附试验检测3株单抗的效价、特异性和相对亲和力，并对其血清学反应进行分析。部分结核病患者的抗原水平明显高于健康对照组。目前可用的检测系统的灵敏度和特异度可能会通过将这些单抗与其他已在使用的单抗结合使用而得到提高。
The secreted Mycobacterium tuberculosis (MTB) proteins, Ag85B and Hsp16.3, have been the focus of intensive research in recent years. These proteins have high sensitivity in bacterium-negative tuberculosis (TB) patients, and are valuable for the rapid diagnosis of bacterium-negative TB. Fusion proteins including multiple antigens such as Ag85B and Hsp16.3 provide improved sensitivity and specificity for serological diagnosis of active TB compared with a single antigen. Many studies have shown that the production of MAbs recognizing a specific repertoire of M. tuberculosis antigens and the tests based on monoclonal antibodies have been found to be valuable in positive detection of TB, particularly for smear-positive pulmonary TB. A number of MAbs are currently used for serodiagnosis of TB. Therefore, an Ag85B-Hsp16.3 fusion protein was expressed and purified using an E. coli system in this study. Three Ag85B-Hsp16.3 fusion protein-specific MAbs were generated by routine murine hybridoma techniques. The titer, specificity, and relative affinity of all three MAbs were determined by ELISA and the serological responses were analyzed. The levels of antigens in a proportion of TB patients were shown to be significantly higher than those in healthy controls. The sensitivity and specificity of the currently available detection systems is likely to be improved by the employment of a combination of these MAbs with others that are already in use.