Directed evolution of an anti-carcinoembryonic antigen scFv with a 4-day monovalent dissociation half-time at 37°C
Directed evolution of an anti-carcinoembryonic antigen scFv with a 4-day monovalent dissociation half-time at 37°C
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DOI：
10.1093/protein/gzh038
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发表时间：
2004-04-01
影响因子：
2.4
通讯作者：
Wittrup, KD
中科院分区：
文献类型：
--
作者：
Graff, CP;Chester, K;Wittrup, KD
已经将scFv工程化以结合癌胚抗原（CEA），在37 ° C下解离半衰期>4天。通过流式细胞术筛选酵母表面展示突变体文库，分离出导致这种亲和力增加的两个突变。通过筛选突变体文库以提高酵母表面展示水平，突变体scFv在酵母分泌系统中的可溶性表达增加了100倍。这种scFv将是有用的，作为一个限制性的情况下，评估的意义，在肿瘤靶向非内化抗原的亲和力。
An scFv has been engineered to bind carcinoembryonic antigen (CEA) with a dissociation half-time >4 days at 37degreesC. Two mutations responsible for this affinity increase were isolated by screening yeast surface-displayed mutant libraries by flow cytometry. Soluble expression of the mutant scFv in a yeast secretion system was increased 100-fold by screening mutant libraries for improved yeast surface display level. This scFv will be useful as a limiting case for evaluating the significance of affinity in tumor targeting to noninternalizing antigens.