Wang Junxia, Tan Yuyu, Ling Jiajun, Zhang Mingxia, Li Liang, Liu Wenli, Huang Mengjiao, Song Jia, Li Ao, Song Yanling, Yang Chaoyong, Zhu Zhi
The MOE Key Laboratory of Spectrochemical Analysis & Instrumentation, The Key Laboratory of Chemical Biology of Fujian Province, State Key Laboratory of Physical Chemistry of Solid Surfaces, Collaborative Innovation Center of Chemistry for Energy Materials, Department of Chemical Biology, Department of Chemical Engineering, College of Chemistry and Chemical Engineering, Xiamen University, Xiamen 361005, China.
Lab Chip. 2021 Mar 21;21(6):1175-1184. doi: 10.1039/d0lc01146k. Epub 2021 Feb 8.
Based on the linkage of genotype and phenotype, display technology has been widely used to generate specific ligands for profiling, imaging, diagnosis and therapy applications. However, due to the lack of effective monoclonal manipulation and affinity evaluation methods, traditional display technology has to undergo tedious steps of selection, clone isolation, amplification, sequencing, synthesis and characterization to obtain the binding sequences. To directly acquire high-affinity clones, we propose a double monoclonal display approach (dm-Display) for peptide screening based on highly paralleled monoclonal manipulation in emulsion droplets. dm-Display can monoclonally link the genotype, phenotype and affinity to realize integrated monoclonal separation, amplification, recognition and staining in one droplet so that discrete high-affinity clones can be quickly extracted. Monoclonal manipulations highly-parallelly occur in millions of droplets so that molecular screening of a highly diverse phage library is achieved. We have screened specific peptide ligands against CD71 and GPC1, proving the feasibility and generality of dm-Display. As a highly efficient ligand screening platform, dm-Display will promote the further development of molecular screening.
基于基因型和表型的关联，展示技术已被广泛用于生成用于分析、成像、诊断和治疗应用的特异性配体。然而，由于缺乏有效的单克隆操作和亲和力评估方法，传统展示技术必须经过繁琐的筛选、克隆分离、扩增、测序、合成和表征步骤才能获得结合序列。为了直接获得高亲和力克隆，我们提出了一种基于乳液滴中高度平行单克隆操作的双单克隆展示方法（dm-Display）用于肽筛选。dm-Display可以将基因型、表型和亲和力进行单克隆连接，从而在一个液滴中实现单克隆分离、扩增、识别和染色，以便快速提取离散的高亲和力克隆。单克隆操作在数百万个液滴中高度平行地进行，从而实现对高度多样化噬菌体文库的分子筛选。我们已经筛选出了针对CD71和GPC1的特异性肽配体，证明了dm-Display的可行性和通用性。作为一个高效的配体筛选平台，dm-Display将推动分子筛选的进一步发展。
