Ho Mitchell, Pastan Ira
Laboratory of Molecular Biology, National Cancer Institute, Bethesda, MD 20892-4264, USA.
Methods Mol Biol. 2009;562:99-113. doi: 10.1007/978-1-60327-302-2_8.
We describe a human cell display strategy to isolate high-affinity single-chain antibody fragments (scFvs) specific for CD22 for the treatment of B-cell malignancies. Our strategy uses flow cytometry and human embryonic kidney 293T (HEK-293T) cells that are widely used for transient protein expression. Flow cytometry enhances the screen's sensitivity thereby allowing us to isolate high-affinity scFvs. Using human cell display, one could isolate and engineer scFvs, single domains, Fabs, or whole IgGs for increased affinity and other biological functions.
我们描述了一种人类细胞展示策略,用于分离针对CD22的高亲和力单链抗体片段(scFv),以治疗B细胞恶性肿瘤。我们的策略使用广泛用于瞬时蛋白表达的流式细胞术和人胚肾293T(HEK-293T)细胞。流式细胞术提高了筛选的灵敏度,从而使我们能够分离高亲和力的scFv。使用人类细胞展示,可以分离和改造scFv、单域抗体、Fab片段或完整的IgG,以提高亲和力和其他生物学功能。