Syngeneic homograft of framework regions enhances the affinity of the mouse anti-human epidermal receptor 2 single-chain antibody e23sFv.
Syngeneic homograft of framework regions enhances the affinity of the mouse anti-human epidermal receptor 2 single-chain antibody e23sFv.
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框架区同基因同种移植增强小鼠抗人表皮受体2单链抗体e23sFv的亲和力
DOI：
10.3892/etm.2020.9568
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发表时间：
2021-03
影响因子：
2.7
通讯作者：
Zhao J
中科院分区：
文献类型：
--
作者：
Ou-Yang Q;Ren JL;Yan B;Feng JN;Yang AG;Zhao J
e23 sFv是一种靶向HER 2的单链可变片段（scFv），在我们之前的研究中被表征为靶向HER 2的肿瘤促凋亡分子的靶向部分。体外抗体亲和力成熟是通过互补决定区（CDR）诱变或框架区（FR）移植来增强抗体亲和力的方法。在本研究中，使用两种策略增强e23 sFv的亲和力。在一种方法中，将定点突变引入e23 sFv的FR（命名为EMEY），而在另一种方法中，用来自最同源的筛选抗体的FR（命名为EX 1和EX 2）取代e23 sFv FR。值得注意的是，与e23 sFv相比，源自FR植入策略的EXl显示出对HER 2的4倍更高的亲和力，并且被内化到HER 2过表达细胞中;然而，与EXl相比，EMEY和EX 2表现出对HER 2的亲和力降低和内化潜力降低。使用分子同源建模和对接获得EX 1和HER 2-EX 1复合物的3D结构，并预测EX 1的HER 2表位和EX 1-HER 2复合物的分子相互作用能。在本研究中，证明了基于序列比对的scFv亲和力提高是可行的和有效的。与定点突变相比，FR嫁接策略更有效、更简便地提高了e23 sFv的亲和力。总之，表明亲和力改善的候选物EX 1可能具有诊断和治疗HER 2过表达肿瘤的巨大潜力。
e23sFv is a HER2-targeted single-chain variable fragment (scFV) that was characterized as the targeting portion of a HER2-targeted tumour proapoptotic molecule in our previous study. In vitro antibody affinity maturation is a method to enhance antibody affinity either by complementarity-determining region (CDR) mutagenesis or by framework region (FR) engraftment. In the present study, the affinity of e23sFv was enhanced using two strategies. In one approach, site-directed mutations were introduced into the FRs of e23sFv (designated EMEY), and in the other approach e23sFv FRs were substituted with FRs from the most homologous screened antibodies (designated EX1 and EX2). Notably, EX1 derived from the FR engraftment strategy demonstrated a 4-fold higher affinity for HER2 compared with e23sFv and was internalized into HER2-overexpressing cells; however, EMEY and EX2 exhibited reduced affinity for HER2 and decreased internalization potential compared with EX1. The 3D structure of EX1 and the HER2-EX1 complex was acquired using molecular homology modelling and docking and the HER2 epitopes of EX1 and the molecular interaction energy of the EX1-HER2 complex were predicted. In the present study, it was demonstrated that scFv affinity improvement based on sequence alignment was feasible and effective. Moreover, the FR grafting strategy was indicated to be more effective and simple compared with site-directed mutagenesis to improve e23sFv affinity. In conclusion, it was indicated that the affinity-improved candidate EX1 may present a great potential for the diagnosis and treatment of HER2-overexpressing tumours.
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影响因子：
64.5
作者：
Briney, Bryan;Sok, Devin;Jardine, Joseph G.;Kulp, Daniel W.;Skog, Patrick;Menis, Sergey;Jacak, Ronald;Kalyuzhniy, Oleksandr;de Val, Natalia;Sesterhenn, Fabian;Le, Khoa M.;Ramos, Alejandra;Jones, Meaghan;Saye-Francisco, Karen L.;Blane, Tanya R.;Spencer, Skye;Georgeson, Erik;Hu, Xiaozhen;Ozorowski, Gabriel;Adachi, Yumiko;Kubitz, Michael;Sarkar, Anita;Wilson, Ian A.;Ward, Andrew B.;Nemazee, David;Burton, Dennis R.;Schief, William R.
通讯作者：
Schief, William R.
影响因子：
5.6
作者：
CHOTHIA, C;LESK, AM
通讯作者：
LESK, AM
DOI：
10.1007/978-1-4939-7447-4_28
发表时间：
2018-01-01
期刊：
PHAGE DISPLAY
影响因子：
--
作者：
Moreira, Gustavo Marcal Schmidt Garcia;Fuehner, Viola;Hust, Michael
通讯作者：
Hust, Michael
影响因子：
4.4
作者：
Wright, GJ;Cherwinski, H;Barclay, AN
通讯作者：
Barclay, AN
影响因子：
5.3
作者：
Teplyakov, Alexey;Obmolova, Galina;Gilliland, Gary L.
通讯作者：
Gilliland, Gary L.