Efficient method to optimize antibodies using avian leukosis virus display and eukaryotic cells.
Efficient method to optimize antibodies using avian leukosis virus display and eukaryotic cells.
复制标题
使用禽白血病病毒展示和真核细胞优化抗体的有效方法。
DOI：
10.1073/pnas.1414754112
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发表时间：
2015
影响因子：
11.1
通讯作者：
Federspiel,MarkJ
中科院分区：
文献类型：
--
作者：
Yu,Changming;Pike,GennettM;Rinkoski,TommyA;Correia,Cristina;Kaufmann,ScottH;Federspiel,MarkJ
基于抗体的疗法现在已经在临床上取得了成功。抗体对靶配体的亲和力和特异性决定了治疗递送的特异性和脱靶副作用。发现和优化的高亲和力抗体的重要的治疗目标可以显着提高了一个强大的，真核展示技术可比噬菌体展示，将克服微生物的蛋白质翻译的限制。真核细胞的使用将改善展示抗体的多样性，所述抗体可以被筛选和优化，以及更无缝地过渡到用于临床生产的大规模哺乳动物表达系统中。在这项研究中，我们证明了真核逆转录病毒，禽白血病病毒（ALV）的复制和多肽展示特性，提供了一个强大的，真核版本的噬菌体展示。通过使用ALV展示优化模型单链Fv抗体的结合亲和力，从微摩尔到皮摩尔水平提高亲和力> 2，000倍。我们相信ALV展示为微生物上的抗体展示提供了延伸，并在真核表达系统中提供了病毒和细胞展示平台。ALV展示应该能够改善适当加工和功能性抗体变体的多样性，所述抗体变体可以被筛选和亲和力优化以改善有希望的抗体候选物。
Antibody-based therapeutics have now had success in the clinic. The affinity and specificity of the antibody for the target ligand determines the specificity of therapeutic delivery and off-target side effects. The discovery and optimization of high-affinity antibodies to important therapeutic targets could be significantly improved by the availability of a robust, eukaryotic display technology comparable to phage display that would overcome the protein translation limitations of microorganisms. The use of eukaryotic cells would improve the diversity of the displayed antibodies that can be screened and optimized as well as more seamlessly transition into a large-scale mammalian expression system for clinical production. In this study, we demonstrate that the replication and polypeptide display characteristics of a eukaryotic retrovirus, avian leukosis virus (ALV), offers a robust, eukaryotic version of bacteriophage display. The binding affinity of a model single-chain Fv antibody was optimized by using ALV display, improving affinity >2,000-fold, from micromolar to picomolar levels. We believe ALV display provides an extension to antibody display on microorganisms and offers virus and cell display platforms in a eukaryotic expression system. ALV display should enable an improvement in the diversity of properly processed and functional antibody variants that can be screened and affinity-optimized to improve promising antibody candidates.