Rapid selection of cell subpopulation-specific human monoclonal antibodies from a synthetic phage antibody library.
Peripheral blood leukocytes incubated with a semisynthetic phage antibody library and fluorochrome-labeled CD3 and CD20 antibodies were used to isolate human single-chain Fv antibodies specific for subsets of blood leukocytes by flow cytometry. Isolated phage antibodies showed exclusive binding to the subpopulation used for selection or displayed additional binding to a restricted population of other cells in the mixture. At least two phage antibodies appeared to display hitherto-unknown staining patterns of B-lineage cells. This approach provides a subtractive procedure to rapidly obtain human antibodies against known and novel surface antigens in their native configuration, expressed on phenotypically defined subpopulations of cells. This approach does not depend on immunization procedures or the necessity to repeatedly construct phage antibody libraries.
de Kruif
J
J
Department of Immunology, Utrecht University, The Netherlands.
Terstappen
L
L
Boel
E
E
Logtenberg
T
T
D000022:Abortion, Spontaneous; D000595:Amino Acid Sequence; D000911:Antibodies, Monoclonal / Q000276:immunology*; D000914:Antibodies, Viral / Q000276:immunology*; D015703:Antigens, CD / Q000276:immunology; D018951:Antigens, CD20; D000944:Antigens, Differentiation, B-Lymphocyte / Q000276:immunology; D001435:Bacteriophages / Q000276:immunology*; D001853:Bone Marrow / Q000276:immunology; D017252:CD3 Complex / Q000276:immunology; D005260:Female; D005333:Fetus; D005434:Flow Cytometry; D006412:Hematopoietic Stem Cells / Q000276:immunology; D006801:Humans; D007128:Immunoglobulin Fragments / Q000276:immunology*; D007143:Immunoglobulin Heavy Chains / Q000737:chemistry / Q000276:immunology; D007147:Immunoglobulin Light Chains / Q000737:chemistry / Q000276:immunology; D007135:Immunoglobulin Variable Region / Q000737:chemistry / Q000276:immunology; D016131:Lymphocyte Subsets / Q000276:immunology*; D008969:Molecular Sequence Data; D011247:Pregnancy
D000911:Antibodies, Monoclonal; D000914:Antibodies, Viral; D015703:Antigens, CD; D018951:Antigens, CD20; D000944:Antigens, Differentiation, B-Lymphocyte; D017252:CD3 Complex; D007128:Immunoglobulin Fragments; D007143:Immunoglobulin Heavy Chains; D007147:Immunoglobulin Light Chains; D007135:Immunoglobulin Variable Region