核仁素
抗体依赖性细胞介导的细胞毒性
抗体
三阴性乳腺癌
癌症研究
细胞毒性
免疫疗法
乳腺癌
癌症
生物
免疫学
化学
单克隆抗体
体外
免疫系统
细胞生物学
生物化学
核仁
细胞质
遗传学
作者
Rita Ribeiro,Jorge M. B. Vítor,Anastasiya Voronovska,João Nuno Moreira,João Gonçalves
标识
DOI:10.1002/biot.202400380
摘要
ABSTRACT Triple‐negative breast cancer (TNBC) is a clinically aggressive subtype of breast cancer that remains an unmet medical need. Because TNBC cells do not express the most common markers of breast cancers, there is an active search for novel molecular targets in triple‐negative tumors. Additionally, this subtype of breast cancer presents strong immunogenic characteristics which have been encouraging the development of immunotherapeutic approaches against the disease. In this context, nucleolin arises as a promising target for immunotherapy against TNBC. Our group has previously developed an anti‐nucleolin VHH‐Fc antibody capable of eliciting antibody‐dependent cellular cytotoxicity (ADCC). Moreover, we constructed and characterized an antibody library, that was screened against nucleolin‐overexpressing cells, originating an enriched anti‐nucleolin antibody pool. In this work, a strategy to select individual clones from the pool was designed, combining NGS data with 3D modeling. Two antibodies demonstrated a significant 4.4‐ and 6.1‐fold increase in binding to nucleolin‐overexpressing and TNBC cells, and an improvement in affinity to the sub‐micromolar range (0.19 µM and 83.69 nM). Additionally, an increment in 4.6‐ and 3.1‐fold in ADCC activity against respective cell lines was observed for the M2 antibody clone. Herein, the affinity maturation strategy developed was validated and corroborated a positive, but not proportional, correlation between antibody binding, affinity, and ADCC.
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