Phage-display selection on tumor histological specimens with laser capture microdissection.
A method was developed to obtain phage-display ligands that bind to a select population of cells in histological specimens of freshly harvested solid human cancers. It combines phage-display panning with laser capture microdissection (LCM). This method allows selection of phage ligands bound to subpopulations of specific cells contained in tumor tissue on histological sections. Naïve phage scFv library was incubated directly on a histological section of human breast cancer that was snap frozen immediately after surgical resection. Tumor and stromal cells were captured by LCM and bound phages were recovered by bacterial infection. Individual phage clones selected after panning were evaluated for their binding ability by immunofluorescence staining on tumor tissue from the same patient. One phage-display antibody clone selected on tumor stroma showed selective binding on tumor stroma but did not bind to malignant cell population. The expressed scFv of this clone showed no significant binding to normal tissue, or 13 other breast cancers, or 4 colon cancer samples. Using the same method, phage display antibody clones were selected on tumor cells which showed binding to tumor cells and normal tissue. This method is applicable for selection of ligands to virtually any portion of a histological specimen amenable to LCM. This may speed the process of generating ligands to any subset of cells or noncellular feature present on histological specimens.
Sun
Yujing
Y
Department of Surgery, Vermont Comprehensive Cancer Center, University of Vermont College of Medicine, Burlington, VT 05405, USA.
Shukla
Girja S
GS
Weaver
Donald
D
Pero
Stephanie C
SC
Krag
David N
DN
D000951:Antigens, Neoplasm / Q000032:analysis*; D001666:Binding Sites, Antibody; D001667:Binding, Competitive; D001943:Breast Neoplasms / Q000276:immunology* / Q000473:pathology; D005260:Female; D006801:Humans; D007135:Immunoglobulin Variable Region / Q000235:genetics / Q000276:immunology*; D007150:Immunohistochemistry; D007834:Lasers; D008024:Ligands; D042282:Microdissection / Q000295:instrumentation*; D008856:Microscopy, Fluorescence; D016612:Paraffin Embedding; D019151:Peptide Library; D017154:Stromal Cells / Q000276:immunology* / Q000473:pathology
D000951:Antigens, Neoplasm; D007135:Immunoglobulin Variable Region; D008024:Ligands; D019151:Peptide Library