Structural insights into humanization of anti-tissue factor antibody 10H10
Structural insights into humanization of anti-tissue factor antibody 10H10
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DOI：
10.1080/19420862.2017.1412026
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发表时间：
2018-01-01
期刊：
影响因子：
5.3
通讯作者：
Gilliland, Gary L.
中科院分区：
文献类型：
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作者：
Teplyakov, Alexey;Obmolova, Galina;Gilliland, Gary L.
针对人组织因子产生的鼠抗体 10H10 的独特之处在于它阻断信号传导途径，从而抑制血管生成和肿瘤生长而不干扰凝血。作为一种潜在的治疗方法，该抗体通过两步程序进行人源化。将抗原结合环移植到选定的人框架上，并使用噬菌体展示文库对所得嵌合抗体进行亲和力成熟。通过人源化变体与亲本小鼠抗体的结构比较，从结构角度分析人源化结果。该分析揭示了框架区中的几个热点似乎影响抗原结合，因此在人类种系选择中应予以考虑。此外，游标区中的一些位置，例如传统上被认为至关重要的重链中的残基 71，似乎能够耐受氨基酸取代，而不会对结合产生任何影响。根据 Kabat 和 Martin 定义的差异，使用短形式和长形式的互补决定区 (CDR) H2 产生了几种人源化变体。此类对的比较表明具有短 CDR H2 的变体始终具有较高的热稳定性。对与结构相关的结合数据的分析指出，ImMunoGeneTics 信息系统 (R) 种系 IGHV1-2*01 是可疑的，因为与同一种系的其他等位基因和其他人类种系相比，有两个潜在的不稳定突变。
Murine antibody 10H10 raised against human tissue factor is unique in that it blocks the signaling pathway, and thus inhibits angiogenesis and tumor growth without interfering with coagulation. As a potential therapeutic, the antibody was humanized in a two-step procedure. Antigen-binding loops were grafted onto selected human frameworks and the resulting chimeric antibody was subjected to affinity maturation by using phage display libraries. The results of humanization were analyzed from the structural perspective through comparison of the structure of a humanized variant with the parental mouse antibody. This analysis revealed several hot spots in the framework region that appear to affect antigen binding, and therefore should be considered in human germline selection. In addition, some positions in the Vernier zone, e.g., residue 71 in the heavy chain, that are traditionally thought to be crucial appear to tolerate amino acid substitutions without any effect on binding. Several humanized variants were produced using both short and long forms of complementarity-determining region (CDR) H2 following the difference in the Kabat and Martin definitions. Comparison of such pairs indicated consistently higher thermostability of the variants with short CDR H2. Analysis of the binding data in relation to the structures singled out the ImMunoGeneTics information system (R) germline IGHV1-2*01 as dubious owing to two potentially destabilizing mutations as compared to the other alleles of the same germline and to other human germlines.