Going native: Direct high throughput screening of secreted full-length IgG antibodies against cell membrane proteins.
Going native: Direct high throughput screening of secreted full-length IgG antibodies against cell membrane proteins.
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DOI：
10.1080/19420862.2017.1381812
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发表时间：
2017
期刊：
影响因子：
5.3
通讯作者：
Griswold KE
中科院分区：
文献类型：
--
作者：
Fang Y;Chu TH;Ackerman ME;Griswold KE
凝胶微滴-荧光激活细胞分选（GMD-FACS）是用于重组蛋白文库的创新的高通量筛选平台，并且我们在这里显示GMD-FACS可以克服与用于抗体文库的常规筛选方法相关的许多限制。例如，噬菌体和细胞表面展示受益于异常高的通量，但通常需要高质量的可溶性抗原靶标，并且需要使用锚定的抗体片段。相比之下，GMD-FACS测定可以以每秒数千个克隆的速率筛选可溶性、分泌性、全长IgG，并且该技术能够在其天然细胞环境中针对膜蛋白靶进行直接筛选。在概念验证实验中，只需三天即可从过量10，000倍的抗CCR 5克隆中高效富集罕见的抗EGFR抗体克隆。展望未来，GMD-FACS有可能有助于抗体的发现和困难的目标，如离子通道和G蛋白偶联受体的工程。
Gel microdroplet – fluorescence activated cell sorting (GMD-FACS) is an innovative high throughput screening platform for recombinant protein libraries, and we show here that GMD-FACS can overcome many of the limitations associated with conventional screening methods for antibody libraries. For example, phage and cell surface display benefit from exceptionally high throughput, but generally require high quality, soluble antigen target and necessitate the use of anchored antibody fragments. In contrast, the GMD-FACS assay can screen for soluble, secreted, full-length IgGs at rates of several thousand clones per second, and the technique enables direct screening against membrane protein targets in their native cellular context. In proof-of-concept experiments, rare anti-EGFR antibody clones were efficiently enriched from a 10,000-fold excess of anti-CCR5 clones in just three days. Looking forward, GMD-FACS has the potential to contribute to antibody discovery and engineering for difficult targets, such as ion channels and G protein-coupled receptors.
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影响因子：
56.9
作者：
HUSE, WD;SASTRY, L;LERNER, RA
通讯作者：
LERNER, RA
影响因子：
5.3
作者：
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影响因子：
5.3
作者：
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通讯作者：
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影响因子：
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作者：
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通讯作者：
Cochran JR
影响因子：
5.7
作者：
Kearns, Jeffrey D.;Bukhalid, Raghida;Wolf, Beni B.
通讯作者：
Wolf, Beni B.