Using phage and yeast display to select hundreds of monoclonal antibodies: application to antigen 85, a tuberculosis biomarker.
Using phage and yeast display to select hundreds of monoclonal antibodies: application to antigen 85, a tuberculosis biomarker.
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使用噬菌体和酵母展示来选择数百种单克隆抗体：抗原85（结核病生物标志物）的应用。
DOI：
10.1371/journal.pone.0049535
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发表时间：
2012
期刊：
影响因子：
3.7
通讯作者：
Bradbury AR
中科院分区：
文献类型：
--
作者：
Ferrara F;Naranjo LA;Kumar S;Gaiotto T;Mukundan H;Swanson B;Bradbury AR
结核病 (TB) 是一项重大的全球健康挑战，每年夺去近 200 万人的生命，目前的诊断方法既耗时又不充分。在感染过程中，许多在感染过程中发挥作用的细菌分子被释放，并被提议作为早期结核病诊断的生物标志物。抗原 85 (Ag85) 是最丰富的分泌性结核病蛋白，也是该诊断方法的潜在靶点。直接检测此类细菌靶标的瓶颈之一是能否获得稳定、灵敏、特异性的抗体。使用 Ag85 作为模型，我们描述了一种使用噬菌体和酵母展示的新型组合来选择针对任何潜在靶标的抗体的方法，该组合利用了每种方法的优点。初步筛选中鉴定出的 111 种特异性抗体证明了这种方法的有效性。在夹心测定中评估它们与不同 Ag85 亚基的结合、亲和力和活性。这种方法的新颖性在于可以通过酵母展示在单个实验中筛选噬菌体抗体选择的全部输出。这可以被认为类似于进行一百万次 ELISA。以这种方式鉴定的单克隆抗体 (mAb) 对抗原表现出高结合亲和力和选择性，并且比通过相对昂贵且耗时的技术生产的传统 mAb 具有优势。该方法具有广泛的适用性，如果需要，可以显着提高所选抗体的亲和力。
Current diagnostic methods for tuberculosis (TB), a major global health challenge that kills nearly two million people annually, are time-consuming and inadequate. During infection a number of bacterial molecules that play a role in the infective process are released and have been proposed as biomarkers for early TB diagnosis. Antigen 85 (Ag85) is the most abundant secreted TB protein, and a potential target for this diagnostic approach. One of the bottlenecks in the direct detection of such bacterial targets is the availability of robust, sensitive, specific antibodies. Using Ag85 as a model, we describe a method to select antibodies against any potential target using a novel combination of phage and yeast display that exploits the advantage of each approach. The efficiency of this approach was attested to by the 111 specific antibodies identified in initial screens. These were assessed for binding to the different Ag85 subunits, affinity, and activity in sandwich assays. The novelty of this approach lies in the possibility of screening the entire output of a phage antibody selection in a single experiment by yeast display. This can be considered analogous to carrying out a million ELISAs. The monoclonal antibodies (mAbs) identified in this way show high binding affinity and selectivity for the antigens and offer an advantage over traditional mAbs produced by relatively expensive and time consuming techniques. This approach has wide applicability, and the affinity of selected antibodies can be significantly improved, if required.
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基于检测抗原85复合物的间接ELISA方案，印度人群中结核病的诊断：一项前瞻性队列研究。
影响因子：
3.7
作者：
Kashyap, Rajpal S.;Rajan, Anju N.;Ramteke, Sonali S.;Agrawal, Vijay S.;Kelkar, Sanjivani S.;Purohit, Hemant J.;Taori, Girdhar M.;Daginawala, Hatim F.
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Daginawala, Hatim F.
影响因子：
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影响因子：
6.8
作者：
Gai, S. Annie;Wittrup, K. Dane
通讯作者：
Wittrup, K. Dane
影响因子：
56.9
作者：
Bostrom, Jenny;Yu, Shang-Fan;Fuh, Germaine
通讯作者：
Fuh, Germaine
影响因子：
3.1
作者：
Bentley-Hibbert, SI;Quan, X;Godfrey, HP
通讯作者：
Godfrey, HP