Selection and affinity maturation of human antibodies against rabies virus from a scFv gene library using ribosome display.
Selection and affinity maturation of human antibodies against rabies virus from a scFv gene library using ribosome display.
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DOI：
10.1016/j.jbiotec.2009.09.022
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发表时间：
2009-12
影响因子：
4.1
通讯作者：
Xiao-ling Zhao;Wei-qiang Chen;Zhi-Hua Yang;Jing-mei Li;Su-juan Zhang;L. Tian
中科院分区：
文献类型：
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作者：
Xiao-ling Zhao;Wei-qiang Chen;Zhi-Hua Yang;Jing-mei Li;Su-juan Zhang;L. Tian
利用核糖体展示技术从人源抗狂犬病病毒单链抗体基因库中筛选出有用的抗体片段。以重组狂犬病毒糖蛋白（RVGp）为抗原，分离特异性单链抗体。在五轮选择后，分析表明scFv-核糖体-mRNA复合物针对RVGp被特异性选择。序列分析表明，突变是随机引入的PCR轮之间的选择和高亲和力的变异体分离。所获得的单链抗体具有高亲和力，能特异性识别RVGp，并显示出与狂犬病病毒的结合活性。这些scFv有可能包含在旨在应用于狂犬病暴露后预防的几种人单克隆抗体（MAb）的组合中。核糖体展示技术是快速分离人抗体片段的有力工具，在体外亲和力成熟和分子进化方面具有卓越的优势。
We selected useful antibody fragments against rabies virus from a human single chain variable fragment (scFv) gene library using ribosome display technique. The recombinant rabies virus glycoprotein (RVGp) was used as an antigen to isolate specific scFvs. After five rounds of selection, the analysis demonstrated that scFv–ribosome–mRNA complexes were specifically selected against RVGp. Sequence analysis showed that mutations were introduced at random by PCR among the rounds of selection and variants with high affinity were isolated. The obtained scFvs with high affinity could recognize RVGp specifically and showed binding activity to rabies virus. These scFvs were potential for inclusion in a combination of several human monoclonal antibodies (MAbs) aimed for application in rabies post-exposure prophylaxis. Ribosome display technology is a robust tool for rapid isolation of human antibody fragments, and has exceptional strength in affinity maturation and molecular evolution in vitro.