title: Selection of a breast cancer subpopulation-specific antibody using phage display on tissue sections.
abstract: Breast cancer tumors are composed of heterogeneous cell populations. These populations display a high variance in morphology, growth and metastatic propensity. They respond differently to therapeutic interventions, and some may be more prone to cause recurrence. Studying individual subpopulations of breast cancer may provide crucial knowledge for the development of individualized therapy. However, this process is challenged by the availability of biomarkers able to identify subpopulations specifically. Here, we demonstrate an approach for phage display selection of recombinant antibody fragments on cryostat sections of human breast cancer tissue. This method allows for selection of recombinant antibodies binding to antigens specifically expressed in a small part of the tissue section. In this case, a CD271(+) subpopulation of breast cancer cells was targeted, and these may be potential breast cancer stem cells. We isolated an antibody fragment LH 7, which in immunohistochemistry experiments demonstrates specific binding to breast cancer subpopulations. The selection of antibody fragments directly on small defined areas within a larger section of malignant tissue is a novel approach by which it is possible to better target cellular heterogeneity in proteomic studies. The identification of novel biomarkers is relevant for our understanding and intervention in human diseases. The selection of the breast cancer-specific antibody fragment LH 7 may reveal novel subpopulation-specific biomarkers, which has the potential to provide new insight and treatment strategies for breast cancer.
authors.affiliation: Department of Molecular Biology and Genetics, Aarhus University, Aarhus, Denmark.
authors.forename: Simon Asbjørn
authors.initials: SA
authors.lastname: Larsen
authors.forename: Theresa
authors.initials: T
authors.lastname: Meldgaard
authors.forename: Agla J
authors.initials: AJ
authors.lastname: Fridriksdottir
authors.forename: Simon
authors.initials: S
authors.lastname: Lykkemark
authors.forename: Pi Camilla
authors.initials: PC
authors.lastname: Poulsen
authors.forename: Laura F
authors.initials: LF
authors.lastname: Overgaard
authors.forename: Helene Bundgaard
authors.initials: HB
authors.lastname: Petersen
authors.forename: Ole William
authors.initials: OW
authors.lastname: Petersen
authors.forename: Peter
authors.initials: P
authors.lastname: Kristensen
mesh_terms: D000912:Antibodies, Neoplasm / Q000276:immunology; D000918:Antibody Specificity / Q000276:immunology; D000951:Antigens, Neoplasm / Q000276:immunology*; D014408:Biomarkers, Tumor / Q000032:analysis; D001943:Breast Neoplasms / Q000235:genetics / Q000276:immunology*; D061505:Cell Surface Display Techniques / Q000379:methods*; D002478:Cells, Cultured; D004797:Enzyme-Linked Immunosorbent Assay; D005260:Female; D006801:Humans; D009419:Nerve Tissue Proteins / Q000276:immunology*; D017475:Receptors, Nerve Growth Factor / Q000276:immunology*; D061905:Single-Domain Antibodies / Q000276:immunology*; D013194:Staining and Labeling / Q000379:methods
chemical_list: D000912:Antibodies, Neoplasm; D000951:Antigens, Neoplasm; D014408:Biomarkers, Tumor; C484065:NGFR protein, human; D009419:Nerve Tissue Proteins; D017475:Receptors, Nerve Growth Factor; D061905:Single-Domain Antibodies