Targeting ErbB3 Receptor in Cancer with Inhibitory Antibodies from Llama.
Targeting ErbB3 Receptor in Cancer with Inhibitory Antibodies from Llama.
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靶向癌症中的ERBB3受体，其乳腺抑制性抗体。
DOI：
10.3390/biomedicines9091106
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发表时间：
2021-08-28
期刊：
影响因子：
4.7
通讯作者：
Shamova OV
中科院分区：
文献类型：
--
作者：
Eliseev IE;Ukrainskaya VM;Yudenko AN;Mikushina AD;Shmakov SV;Afremova AI;Ekimova VM;Vronskaia AA;Knyazev NA;Shamova OV
人类ErbB3受体对肿瘤中EGFR和HER2受体酪氨酸激酶的药理抑制具有抗性，使其成为重要的治疗靶点。目前正在开发的几种抗erbb3单克隆抗体都是经典的免疫球蛋白。我们采用了不同的方法，通过免疫骆驼抗体库的噬菌体展示，发现了一组新的靶向ErbB3细胞外结构域的重链抗体。我们首先在大肠杆菌中产生三种选定的单域抗体，命名为BCD090-P1， BCD090-M2和BCD090-M456，作为SUMO融合物，每升培养物产生高达180毫克的重组蛋白。然后，我们研究了折叠、聚集和二硫键的形成，并通过最强的候选物BCD090-P1在8 M尿素中发生半变性，证明了它们的最终稳定性。在表面等离子体共振实验中，两种最有效的抗体BCD090-P1和BCD090-M2分别以1.6 nM和15 nM的亲和力结合ErbB3的细胞外结构域进行单价相互作用。通过免疫荧光共聚焦显微镜在四种不同的ErbB3+癌细胞系上证实了受体的结合。我们观察到BCD090-P1和BCD090-M2与受体上的两个不同的表位非竞争性结合。两种抗体均能抑制erbb3驱动的MCF-7乳腺腺癌细胞和her2过表达的SK-BR-3细胞的增殖，EC50在0.1 ~ 25 μg/mL范围内。BCD090-M2直接阻断配体结合，而BCD090-P1不与配体竞争，可能通过不同的变构机制起作用。我们期望这些羊驼抗体可以用于设计新的双异位抗erbb3或双特异性抗erbb2 /3抗体。
The human ErbB3 receptor confers resistance to the pharmacological inhibition of EGFR and HER2 receptor tyrosine kinases in cancer, which makes it an important therapeutic target. Several anti-ErbB3 monoclonal antibodies that are currently being developed are all classical immunoglobulins. We took a different approach and discovered a group of novel heavy-chain antibodies targeting the extracellular domain of ErbB3 via a phage display of an antibody library from immunized llamas. We first produced three selected single-domain antibodies, named BCD090-P1, BCD090-M2, and BCD090-M456, in E. coli, as SUMO fusions that yielded up to 180 mg of recombinant protein per liter of culture. Then, we studied folding, aggregation, and disulfide bond formation, and showed their ultimate stability with half-denaturation of the strongest candidate, BCD090-P1, occurring in 8 M of urea. In surface plasmon resonance experiments, two most potent antibodies, BCD090-P1 and BCD090-M2, bound the extracellular domain of ErbB3 with 1.6 nM and 15 nM affinities for the monovalent interaction, respectively. The receptor binding was demonstrated by immunofluorescent confocal microscopy on four different ErbB3+ cancer cell lines. We observed that BCD090-P1 and BCD090-M2 bind noncompetitively to two distinct epitopes on the receptor. Both antibodies inhibited the ErbB3-driven proliferation of MCF-7 breast adenocarcinoma cells and HER2-overexpressing SK-BR-3 cells, with the EC50 in the range of 0.1–25 μg/mL. BCD090-M2 directly blocks ligand binding, whereas BCD090-P1 does not compete with the ligand and presumably acts through a distinct allosteric mechanism. We anticipate that these llama antibodies can be used to engineer new biparatopic anti-ErbB3 or bispecific anti-ErbB2/3 antibodies.
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影响因子：
3.7
作者：
Göstring L;Malm M;Höidén-Guthenberg I;Frejd FY;Ståhl S;Löfblom J;Gedda L
通讯作者：
Gedda L
影响因子：
4
作者：
Doll, Sophia;Burlingame, Alma L.
通讯作者：
Burlingame, Alma L.
影响因子：
11.4
作者：
BENLEVY, R;PATERSON, HF;YARDEN, Y
通讯作者：
YARDEN, Y
DOI：
10.1073/pnas.91.17.8132
发表时间：
1994-08-16
影响因子：
11.1
作者：
GUY, PM;PLATKO, JV;CARRAWAY, KL
通讯作者：
CARRAWAY, KL
影响因子：
2.4
作者：
Kronqvist, Nina;Malm, Magdalena;Lofblom, John
通讯作者：
Lofblom, John