title: Raising an Antibody Specific to Breast Cancer Subpopulations Using Phage Display on Tissue Sections.
abstract: OBJECTIVE
  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.
  
  
  METHODS
  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.
  
  
  RESULTS
  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.
  
  
  CONCLUSIONS
  Further studies characterizing this antibody fragment, the subpopulation it binds and the cognate antigen may unearth novel biomarkers of clinical relevance.
authors.affiliation: Department of Molecular Biology and Genetics, Aarhus University, Aarhus, Denmark.
authors.forename: Simon Asbjørn
authors.initials: SA
authors.lastname: Larsen
authors.affiliation: Department of Engineering, Aarhus University, Aarhus, Denmark.
authors.forename: Theresa
authors.initials: T
authors.lastname: Meldgaard
authors.affiliation: Department of Cellular and Molecular Medicine, Centre for Biological Disease Analysis and Danish Stem Cell Centre, University of Copenhagen, Copenhagen, Denmark.
authors.forename: Agla Jael Rubner
authors.initials: AJ
authors.lastname: Fridriksdottir
authors.affiliation: Department of Clinical Medicine, Aarhus University, Aarhus, Denmark Sino-Danish Centre for Education and Research (SDC), Aarhus, Denmark.
authors.forename: Simon
authors.initials: S
authors.lastname: Lykkemark
authors.affiliation: Department of Engineering, Aarhus University, Aarhus, Denmark.
authors.forename: Pi Camilla
authors.initials: PC
authors.lastname: Poulsen
authors.affiliation: Department of Engineering, Aarhus University, Aarhus, Denmark.
authors.forename: Laura Falkensteen
authors.initials: LF
authors.lastname: Overgaard
authors.affiliation: Department of Engineering, Aarhus University, Aarhus, Denmark.
authors.forename: Helene Bundgaard
authors.initials: HB
authors.lastname: Petersen
authors.affiliation: Department of Cellular and Molecular Medicine, Centre for Biological Disease Analysis and Danish Stem Cell Centre, University of Copenhagen, Copenhagen, Denmark.
authors.forename: Ole William
authors.initials: OW
authors.lastname: Petersen
authors.affiliation: Department of Engineering, Aarhus University, Aarhus, Denmark pk@eng.au.dk.
authors.forename: Peter
authors.initials: P
authors.lastname: Kristensen
mesh_terms: D000918:Antibody Specificity; D000951:Antigens, Neoplasm / Q000276:immunology; D001435:Bacteriophages / Q000276:immunology*; D014408:Biomarkers, Tumor / Q000276:immunology; D001943:Breast Neoplasms / Q000276:immunology* / Q000473:pathology; D002478:Cells, Cultured; D005260:Female; D006801:Humans; D007150:Immunohistochemistry; D019151:Peptide Library
chemical_list: D000951:Antigens, Neoplasm; D014408:Biomarkers, Tumor; D019151:Peptide Library
keywords: CD271+; Phage display; biomarkers; breast cancer; shadow stick; tissue