Monoclonal antibodies (mAbs) against severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2), which causes COVID-19, are the important tools both for the diagnosis and therapeutics of this infectious disease. The high-performance antibody against spike protein of SARS-CoV-2 is expected to inhibit the binding of viruses to their receptors on the surface of their target cells. In this study, we propose the novel screening method for mAbs against the pathogenic infectious virus using exosome. By this method, the exosome that artificially expresses SARS-CoV-2 spike protein was purified and used as a virus-like vesicle, which could bind to the viral receptor, angiotensin-converting enzyme 2 (ACE2). As a result, seven mAbs that could bind to the spike protein were obtained and six of these clones could strongly inhibit the binding to ACE2 of both the protein corresponding to the receptor binding domain (RBD) and the exosome expressing the spike protein. Interestingly, some of these antibodies seemed to share their epitopes in RBD, suggesting that highly antigenic sites exist in the spike protein. In view of the neutralizing activities on infection, five clones of these antibodies could inhibit the internalization of vesicular stomatitis virus-based pseudo viruses expressing various types of spike proteins derived from SARS-CoV-2 variants. In addition, these antibodies inhibited the infection of SARS-CoV-2 to cultured mammalian cells. These antibodies are expected to be utilized for both diagnosis and therapeutics of COVID-19.