噬菌体展示
选择(遗传算法)
生物传感器
吞吐量
高通量筛选
计算生物学
计算机科学
纳米技术
生物
人工智能
遗传学
材料科学
操作系统
抗体
无线
作者
Phebe De Keyser,Valentina Kalichuk,Thomas Zögg,Alexandre Wohlkönig,Stephan Schenck,Janine D. Brunner,Els Pardon,Jan Steyaert
标识
DOI:10.1101/2024.06.26.600780
摘要
Biopanning methods to select target-specific Nanobodies® (Nbs) involve presenting the antigen, immobilized on plastic plates or magnetic beads, to Nb libraries displayed on phage. Most routines are operator-dependent, labor-intensive and often material- and time-consuming. Here we validate an improved panning strategy that uses biosensors to present the antigen to phage-displayed Nbs in a well. The use of automated Octet biolayer interferometry sensors (Sartorius) enables high throughput and precise control over each step. By playing with association and dissociation times and buffer composition, one can efficiently decrease the background of aspecific and low-affinity Nbs, reducing the rounds of panning needed for the enrichment of high-affinity binders. Octet panning also enables the use of unpurified target proteins and unpurified phage from a bacterial culture supernatant. Additionally, downscaling to a 384-well format significantly reduces the amount of protein required. Moreover, enrichment of binders can be quantified by monitoring phage binding to the target by interferometry, omitting additional phage titration steps. Routinely, up to 3 rounds of Octet panning can be performed in only 5 days to deliver target-specific binders, ready for screening and characterization using the same Octet instrument.
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