Department of Molecular Biology and Genetics, Aarhus University, Aarhus, Denmark.
Department of Engineering, Aarhus University, Aarhus, Denmark.
Cancer Genomics Proteomics. 2016 Jan-Feb;13(1):21-30.
BACKGROUND/AIM: Primary tumors display a great level of intra-tumor heterogeneity in breast cancer. The current lack of prognostic and predictive biomarkers limits accurate stratification and the ability to predict response to therapy. The aim of the present study was to select recombinant antibody fragments specific against breast cancer subpopulations, aiding the discovery of novel biomarkers.
Recombinant antibody fragments were selected by phage display. A novel shadowstick technology enabled the direct selection using tissue sections of antibody fragments specific against small subpopulations of breast cancer cells. Selections were performed against a subpopulation of breast cancer cells expressing CD271+, as these previously have been indicated to be potential breast cancer stem cells. The selected antibody fragments were screened by phage ELISA on both breast cancer and myoepithelial cells. The antibody fragments were validated and evaluated by immunohistochemistry experiments.
Our study revealed an antibody fragment, LH8, specific for breast cancer cells. Immunohistochemistry results indicate that this particular antibody fragment binds an antigen that exhibits differential expression in different breast cancer subpopulations.
Further studies characterizing this antibody fragment, the subpopulation it binds and the cognate antigen may unearth novel biomarkers of clinical relevance.
背景/目的：乳腺癌原发肿瘤存在高度的肿瘤内异质性。目前缺乏预后和预测生物标志物，限制了准确分层和预测治疗反应的能力。本研究旨在选择针对乳腺癌亚群的重组抗体片段，以辅助发现新的生物标志物。
通过噬菌体展示选择重组抗体片段。一种新颖的ShadowStick 技术能够直接使用组织切片对针对乳腺癌细胞小亚群的抗体片段进行选择。选择针对表达 CD271+的乳腺癌细胞亚群进行，因为这些细胞此前已被表明是潜在的乳腺癌干细胞。通过噬菌体 ELISA 对乳腺癌细胞和肌上皮细胞筛选选定的抗体片段。通过免疫组织化学实验对抗体片段进行验证和评估。
我们的研究揭示了一种针对乳腺癌细胞的抗体片段 LH8。免疫组织化学结果表明，该抗体片段结合的抗原在不同的乳腺癌亚群中存在差异表达。
进一步研究该抗体片段、它结合的亚群和相应的抗原，可能会发现具有临床相关性的新的生物标志物。
