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首页> 外文期刊>The Journal of Biochemistry >Construction and characterization of single-chain antibodies against human insulin-like growth factor-I receptor from hybridomas producing 1H7 or 3B7 monoclonal antibody.
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Construction and characterization of single-chain antibodies against human insulin-like growth factor-I receptor from hybridomas producing 1H7 or 3B7 monoclonal antibody.

机译：产生1H7或3B7单克隆抗体的杂交瘤中抗人胰岛素样生长因子-I受体单链抗体的构建和表征。

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摘要

Recombinant antibody consisting of the single-chain variable fragment (scFv) of 1H7 monoclonal antibody against insulin-like growth factor-I receptor (IGF-IR) and human IgG(1) Fc domain, scFv-Fc, has been found to exhibit inhibitory effects on breast cancer growth in vitro and in vivo [Li et al. (2000) Cancer Immunol. Immunother. 49, 243; Sachdev et al. (2003) Cancer Res. 63, 627]. Various types of scFvs from hybridomas producing 1H7 or 3B7 mAb were constructed using conventional phage display technology to further characterize the specificity and affinity of anti-IGF-IR mAbs. Binding studies performed using either phage antibodies or soluble scFv proteins to IGF-IR or insulin receptor (IR) and IGF-IR pre-incubated with mAbs suggested that (i) 1H7 and 3B7 bind to IGF-IR but do not bind to its structurally related IR, (ii) either the VL-VH or VH-VL sequence order does not apparently affect specificity for IGF-IR and (iii) 1H7 and 3B7 bind the independent epitopes, located in or near the N-terminal (440-514) and C-terminal (62-184) domains of the alpha subunit, respectively. This study not only revealed new information on binding regions for two anti-IGF-IR mAbs, but also provided the scFv genes as tools for further manipulation of the affinity or development of new IGF-IR-targeted cancer therapeutics.
机译：已发现由1H7单克隆抗体的单链可变片段（scFv）构成的重组抗体具有抗胰岛素样生长因子-I受体（IGF-IR）和人IgG（1）Fc结构域的重组抗体，具有抑制作用。体外和体内对乳腺癌生长的影响[Li等。 （2000）Cancer Immunol。免疫其他。 49，243; Sachdev等。 （2003）Cancer Res。 63，627]。使用常规噬菌体展示技术构建了来自产生1H7或3B7 mAb的杂交瘤的各种类型的scFv，以进一步表征抗IGF-1R mAb的特异性和亲和力。使用噬菌体抗体或可溶性scFv蛋白与IGF-IR或胰岛素受体（IR）进行的结合研究以及与mAb预孵育的IGF-IR表明（i）1H7和3B7与IGF-IR结合但在结构上不与其结合相关的IR，（ii）VL-VH或VH-VL序列顺序显然不会影响对IGF-IR的特异性，并且（iii）1H7和3B7结合位于N末端或附近的独立表位（440-514） ）和α亚基的C端（62-184）域。这项研究不仅揭示了有关两种抗IGF-1R mAb结合区的新信息，而且还提供了scFv基因作为进一步操纵以IGF-1R为靶标的新型癌症治疗剂的亲和力或开发工具。

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