Generation of a novel affibody molecule targeting Chlamydia trachomatis MOMP.
Generation of a novel affibody molecule targeting Chlamydia trachomatis MOMP.
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靶向沙眼衣原体 MOMP 的新型亲和体分子的生成
DOI：
10.1007/s00253-021-11128-x
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发表时间：
2021-03
影响因子：
5
通讯作者：
Zhu S
中科院分区：
文献类型：
--
作者：
Li M;Shi W;Yang J;Wang Q;Dong H;Chen J;Zhang L;Zhu S
沙眼衣原体（C.沙眼）是全世界可预防失明的主要原因，也是细菌性传播疾病的最普遍原因。目前，没有可用的疫苗，抗生素治疗后复发是实质性问题。主要外膜蛋白（MOMP）占C.沙眼衣原体，作为三聚体孔蛋白起作用，并且它是高度抗原性的。因此，MOMP是衣原体疫苗开发和靶向治疗的最有希望的候选者。亲和体（Affibody）是一类来源于金黄色葡萄球菌A蛋白结合区Z结构域的新型亲和配体，作为一种可行的抗体替代物，已成为研究者关注的焦点。本研究利用噬菌体展示肽库技术筛选出MOMP靶向的噬菌体抗体分子（ZMOMP：461）。进一步通过表面等离子体共振（SPR）和Western blot对亲和性和特异性进行了表征。免疫荧光分析（IFA）显示，MOMP结合抗体能识别感染C.沙眼免疫沉淀试验进一步证实ZMOMP：461分子特异性识别松弛三聚体MOMP上的表位。我们的研究结果提供了强有力的证据表明，抗体分子（ZMOMP：461）作为MOMP抗体的替代物用于生物学应用，并具有很大的潜力，为靶向治疗提供药物。·我们筛选了一种针对沙眼衣原体MOMP的新型亲和体分子ZMOMP：461。· ZMOMP：461以高亲和力和特异性识别重组和天然MOMP。· ZMOMP：461可以内化到活靶细胞中。在线版本包含补充材料，可通过10.1007/s 00253 -021-11128-x获得。
Chlamydia trachomatis (C. trachomatis) is the leading cause of preventable blindness worldwide and the most prevalent cause of bacterial sexually transmitted diseases. At present, there is no available vaccine, and recurrences after antibiotics treatment are substantial problems. Major outer membrane protein (MOMP) accounts for 60% of the outer mass of C. trachomatis, functioning as trimeric porin, and it is highly antigenic. Therefore, MOMP is the most promising candidate for vaccine developing and target therapy of Chlamydia. Affibody, a new class of affinity ligands derived from the Z-domain in the binding region of Staphylococcus aureus protein A, has been the focus of researchers as a viable alternative to antibodies. In this study, the MOMP-targeted affibody molecule (ZMOMP:461) was screened by phage-displayed peptide library. Further, the affinity and specificity were characterized by surface plasmon resonance (SPR) and Western blot. Immunofluorescence assay (IFA) indicated that the MOMP-binding affibody could recognize native MOMP in HeLa229 cells infected C. trachomatis. Immunoprecipitation assay confirmed further that ZMOMP:461 molecule specifically recognizes the epitope on relaxed trimer MOMP. Our findings provide strong evidence that affibody molecule (ZMOMP:461) serves as substitute for MOMP antibody for biological applications and has a great potential for delivering drugs for target therapy. • We screened a novel affibody molecule ZMOMP:461 targeting Chlamydia trachomatis MOMP. • ZMOMP:461 recognizes the recombinant and native MOMP with high affinity and specificity. • ZMOMP:461 could be internalized into live target cells. The online version contains supplementary material available at 10.1007/s00253-021-11128-x.
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DOI：
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期刊：
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