Isolation of osteosarcoma-associated human antibodies from a combinatorial Fab phage display library.
Osteosarcoma, a highly malignant disease, is the most common primary bone tumor and is frequently found in children and adolescents. In order to isolate antibodies against osteosarcoma antigens, a combinatorial osteosarcoma Fab library displayed on the surface of phages was used. After three rounds of selection on the surface of tumor cells, several osteosarcoma-reactive Fabs were detected. From these Fabs, five were better characterized, and despite having differences in their VH (heavy chain variable domain) and Vkappa (kappa chain variable domain) regions, they all bound to a protein with the same molecular mass. Further analysis by cell ELISA and immunocytochemistry suggested that the Fabs recognize a membrane-associated tumor antigen expressed in higher amounts in neoplasic cells than in normal tissue. These results suggest that the human Fabs selected in this work are a valuable tool for the study of this neoplasia.
Dantas-Barbosa
Carmela
C
Institut Gustave Roussy, UPRES EA 3535 PR2. 39, Rue Camille Desmoulins, 94805 Villejuif, France. carmeladantas@hotmail.com
Faria
Fabrícia P
FP
Brigido
Marcelo M
MM
Maranhão
Andrea Q
AQ
D000912:Antibodies, Neoplasm / Q000737:chemistry / Q000276:immunology* / Q000302:isolation & purification*; D002460:Cell Line; D020650:Combinatorial Chemistry Techniques / Q000379:methods; D006801:Humans; D007140:Immunoglobulin Fab Fragments / Q000737:chemistry / Q000276:immunology* / Q000302:isolation & purification; D012516:Osteosarcoma / Q000276:immunology*; D019151:Peptide Library; D014407:Tumor Cells, Cultured
D000912:Antibodies, Neoplasm; D007140:Immunoglobulin Fab Fragments; D019151:Peptide Library