Fransson Johan, Tornberg Ulla-Carin, Borrebaeck Carl A K, Carlsson Roland, Frendéus Björn
Department of Immunotechnology, Lund University, Lund, Sweden.
Int J Cancer. 2006 Jul 15;119(2):349-58. doi: 10.1002/ijc.21829.
Novel panning and screening methodology was devised to isolate high affinity human recombinant scFv antibody fragments with functionally associated properties in B lymphoma cells. The approach was used to generate a panel of apoptosis-inducing antibodies specific for antigens differentially expressed in B lymphoma vs. T leukaemia cells. The selections resulted in an antibody pool with near perfect selectivity (>99%) for the B lymphoma target cells. Randomly picked clones (72) revealed 7 unique antibody genotypes. Six of these rapidly induced apoptosis in target cells. Following the conversion to full IgGs, the antibodies were shown to be specific for HLA-DR/DP, the B-cell receptor mu chain and for CD54/ICAM-1. The latter receptor was not previously associated with apoptotic properties in B-cell lymphomas. Anti-ICAM-1 IgG induced apoptosis in a broad range of B lymphoma cell lines and were shown by immunohistochemistry to bind strongly to B lymphoma tissue obtained from 5 different B lymphoma patients. The recombinant IgG antibodies had affinities in the subnanomolar (0.3 nM) to nanomolar (3 nM) range. The described technology is generally applicable for the rapid isolation of high affinity human antibodies with specificity for differentially expressed cell surface receptors with intrinsic negative or positive signalling properties from naïve phage libraries.
设计了新型淘选和筛选方法，以从B淋巴瘤细胞中分离出具有功能相关特性的高亲和力人重组单链抗体片段。该方法用于生成一组对B淋巴瘤细胞与T白血病细胞中差异表达的抗原具有特异性的凋亡诱导抗体。筛选结果产生了一个对B淋巴瘤靶细胞具有近乎完美选择性（>99%）的抗体库。随机挑选的克隆（72个）显示出7种独特的抗体基因型。其中六种能在靶细胞中迅速诱导凋亡。转化为完整的IgG后，这些抗体被证明对HLA-DR/DP、B细胞受体μ链和CD54/ICAM-1具有特异性。后一种受体以前未与B细胞淋巴瘤的凋亡特性相关联。抗ICAM-1 IgG在多种B淋巴瘤细胞系中诱导凋亡，并通过免疫组织化学显示与从5名不同B淋巴瘤患者获得的B淋巴瘤组织强烈结合。重组IgG抗体的亲和力在亚纳摩尔（0.3 nM）至纳摩尔（3 nM）范围内。所描述的技术通常适用于从原始噬菌体文库中快速分离对具有内在负或正信号特性的差异表达细胞表面受体具有特异性的高亲和力人抗体。
