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Discovery of internalizing antibodies to basal breast cancer cells.
Protein Engineering, Design and Selection ( IF 2.4 ) Pub Date : 2018-01-05 , DOI: 10.1093/protein/gzx063
Yu Zhou 1 , Hao Zou 1 , Christina Yau 2 , Lequn Zhao 1 , Steven C Hall 3 , Daryl C Drummond 4 , Shauna Farr-Jones 1 , John W Park 5 , Christopher C Benz 2 , James D Marks 1
Affiliation  

We present a strategy to discover recombinant monoclonal antibodies (mAbs) to specific cancers and demonstrate this approach using basal subtype breast cancers. A phage antibody library was depleted of antibodies to common cell surface molecules by incubation with luminal breast cancer cell lines, and then selected on a single basal-like breast cancer cell line (MDA-MB-231) for binding associated receptor-mediated endocytosis. Additional profiling against two luminal and four basal-like cell lines revealed 61 unique basal-specific mAbs from a pool of 1440 phage antibodies. The unique mAbs were further screened on nine basal and seven luminal cell lines to identify those with the greatest affinity, specificity, and internalizing capability for basal-like breast cancer cells. Among the internalizing basal-specific mAbs were those recognizing four transmembrane receptors (EphA2, CD44, CD73 and EGFR), identified by immunoprecipitation-mass spectrometry and yeast-displayed antigen screening. Basal-like breast cancer expression of these four receptors was confirmed using a bioinformatic approach, and expression microarray data on 683 intrinsically subtyped primary breast tumors. This overall approach, which sequentially employs phage display antibody library selection, antigen identification and bioinformatic confirmation of antigen expression by cancer subtypes, offers efficient production of high-affinity mAbs with diagnostic and therapeutic utility against specific cancer subtypes.

中文翻译:

发现基础乳腺癌细胞内在化抗体。

我们提出了一种战略,以发现针对特定癌症的重组单克隆抗体(mAb),并证明了使用基础亚型乳腺癌的这种方法。噬菌体抗体文库通过与腔内乳腺癌细胞系孵育而去除了针对常见细胞表面分子的抗体,然后在单个基底样乳腺癌细胞系(MDA-MB-231)上进行筛选,以结合相关的受体介导的内吞作用。针对两个腔和四个基底样细胞系的其他分析揭示了来自1440个噬菌体抗体库的61个独特的基底特异性mAb。在9个基底细胞系和7个管腔细胞系上进一步筛选了独特的mAb,以鉴定对基底样乳腺癌细胞具有最大亲和力,特异性和内在化能力的单克隆抗体。内在化的基础特异性mAb是识别四种跨膜受体(EphA2,CD44,CD73和EGFR)的那些,通过免疫沉淀质谱和酵母展示的抗原筛选鉴定。使用生物信息学方法证实了这四个受体的基础样乳腺癌表达,并且针对683种内在亚型的原发性乳腺肿瘤表达微阵列数据。该总体方法依次采用噬菌体展示抗体库选择,抗原鉴定和癌症亚型抗原表达的生物信息学确认,可提供高亲和力单克隆抗体的有效生产,并具有针对特定癌症亚型的诊断和治疗用途。使用生物信息学方法证实了这四个受体的基础样乳腺癌表达,并且针对683种内在亚型的原发性乳腺肿瘤表达微阵列数据。该总体方法依次采用噬菌体展示抗体库选择,抗原鉴定和癌症亚型抗原表达的生物信息学确认,可提供高亲和力单克隆抗体的有效生产,并具有针对特定癌症亚型的诊断和治疗用途。使用生物信息学方法证实了这四个受体的基础样乳腺癌表达,并且针对683种内在亚型的原发性乳腺肿瘤表达微阵列数据。该总体方法依次采用噬菌体展示抗体库选择,抗原鉴定和癌症亚型抗原表达的生物信息学确认,可提供高亲和力单克隆抗体的有效生产,并具有针对特定癌症亚型的诊断和治疗用途。
更新日期:2019-11-01
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