There is an interest to develop anti-ROBO4 antibodies that can be suitable for the treatment of cancer. The inventors produced new human antibodies directed against ROBO4 were by a
phage display strategy and the antibody selection was performed by using HEK and HEK stably expressing ROBO4 (HEK-ROBO4) cell lines. As a first step, a batch of 6 different
antibodies named D3, H3, H9, E11, H11 and G12 was tested for their binding properties. The
antibodies were then tested for their ability to inhibit the attachment of tumor cells to
osteoblastic cells in monolayers. Finally the inventors showed that the anti-ROBO4 antibodies E11 and G12 inhibits significantly the formation of the tumor spheroid. The present invention thus relates to anti-ROBO4 human monoclonal antibodies and uses thereof for the treatment of cancer.