Genetically-encoded fragment-based discovery (GE-FBD) of glycopeptide ligands with differential selectivity for antibodies related to mycobacterial infections
Genetically-encoded fragment-based discovery (GE-FBD) of glycopeptide ligands with differential selectivity for antibodies related to mycobacterial infections
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DOI：
10.1039/c7ob02783d
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发表时间：
2018-01-14
影响因子：
3.2
通讯作者：
Derda, Ratmir
中科院分区：
文献类型：
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作者：
Chou, Ying;Kitova, Elena N.;Derda, Ratmir
准确识别由结核分枝杆菌引起的结核病（TB）对于全球疾病管理至关重要。护理点血清学检测可改善结核病诊断;然而，现有血清学诊断的特异性是次优的。我们采用基于基因编码片段的发现（GE-FBD）来选择针对分枝杆菌细胞壁组分脂阿拉伯甘露聚糖（LAM）（一种有效抗原）的抗体的配体。GE-FBD使用了108个七肽的噬菌体展示文库，其用LAM的阿拉伯呋喃糖基六糖片段（Ara（6））化学修饰，并且抗LAM抗体CS-35作为诱饵。该选择产生了对CS-35具有增强的亲和力和选择性的糖肽，但对906.4321抗体没有，两者都以相当的亲和力结合Ara（6）。结合发现的配体Ara（6）-ANSSFAP、Ara（6）-DAHATLR和Ara（6）-TTYVVNP的多价测定在CS-35和906.4321之间表现出高达19倍的区分度。单独使用Ara（6）抗原不能区分这些抗体。因此，GE-FBD产生以增强的特异性区分单克隆抗体的配体。这项技术可以促进发展有效的护理点血清学测试分枝杆菌和其他感染。
Accurate identification of tuberculosis (TB), caused by Mycobacterium tuberculosis, is important for global disease management. Point-of-care serological tests may improve TB diagnosis; however, specificities of available serodiagnostics are sub-optimal. We employed genetically encoded fragment-based discovery (GE-FBD) to select ligands for antibodies directed against the mycobacterial cell wall component lipoarabinomannan (LAM), a potent antigen. GE-FBD employed a phage displayed library of 108 heptapeptides, chemically modified with an arabinofuranosyl hexasaccharide fragment of LAM (Ara(6)), and the anti-LAM antibody CS-35 as a bait. The selection gave rise to glycopeptides with an enhanced affinity and selectivity for CS-35 but not for 906.4321 antibody, both of which bind to Ara(6) with a comparable affinity. Multivalent assays incorporating the discovered ligands Ara(6)-ANSSFAP, Ara(6)-DAHATLR and Ara(6)-TTYVVNP exhibited up to 19-fold discrimination between CS-35 and 906.4321. The use of the Ara(6) antigen alone failed to distinguish these antibodies. Thus, GE-FBD gives rise to ligands that differentiate monoclonal antibodies with enhanced specificity. This technology could facilitate the development of effective point-of-care serological tests for mycobacterial and other infections.