Fortunato Ferrara, Adeline Fanni, Andre A R Teixeira, Esteban Molina, Camila Leal-Lopes, Ashley DeAguero, Sara D'Angelo, M Frank Erasmus, Laura Spector, Luis Antonio Rodriguez Carnero, Jianquan Li, Thomas J Pohl, Nikolai Suslov, Klervi Desrumeaux, Conor McMahon, Sagar Kathuria, Andrew R M Bradbury
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引用次数: 0
Abstract
We previously described an in vitro single-chain fragment (scFv) library platform originally designed to generate antibodies with excellent developability properties. The platform design was based on the use of clinical antibodies as scaffolds into which replicated natural complementarity-determining regions purged of sequence liabilities were inserted, and the use of phage and yeast display to carry out antibody selection. In addition to being developable, antibodies generated using our platform were extremely diverse, with most campaigns yielding sub-nanomolar binders. Here, we describe a platform advancement that incorporates Fab phage display followed by single-chain antibody-binding fragment Fab (scFab) yeast display. The scFab single-gene format provides balanced expression of light and heavy chains, with enhanced conversion to IgG, thereby combining the advantages of scFvs and Fabs. A meticulously engineered, quality-controlled Fab phage library was created using design principles similar to those used to create the scFv library. A diverse panel of binding scFabs, with high conversion efficiency to IgG, was isolated against two targets. This study highlights the compatibility of phage and yeast display with a Fab semi-synthetic library design, offering an efficient approach to generate drug-like antibodies directly, facilitating their conversion to potential therapeutic candidates.
我们以前曾描述过一种体外单链片段(scFv)文库平台,该平台最初是为了产生具有优良开发特性的抗体而设计的。该平台设计的基础是使用临床抗体作为支架,在支架上插入复制的天然互补决定区,并清除序列负债,然后使用噬菌体和酵母展示来进行抗体筛选。除了可开发外,利用我们的平台生成的抗体也极为多样化,大多数活动都能产生亚纳莫尔结合剂。在这里,我们描述了一种平台的进步,它结合了 Fab 噬菌体展示和单链抗体结合片段 Fab(scFab)酵母展示。scFab 单基因格式提供了轻链和重链的平衡表达,增强了向 IgG 的转化,从而结合了 scFv 和 Fab 的优势。我们采用与创建 scFv 文库类似的设计原则,创建了一个精心设计、质量受控的 Fab 噬菌体文库。针对两个靶点分离出了多种具有高IgG转化效率的结合scFabs。这项研究强调了噬菌体和酵母展示与 Fab 半合成文库设计的兼容性,为直接生成药物样抗体提供了一种有效的方法,有助于将其转化为潜在的候选治疗药物。
期刊介绍:
mAbs is a multi-disciplinary journal dedicated to the art and science of antibody research and development. The journal has a strong scientific and medical focus, but also strives to serve a broader readership. The articles are thus of interest to scientists, clinical researchers, and physicians, as well as the wider mAb community, including our readers involved in technology transfer, legal issues, investment, strategic planning and the regulation of therapeutics.