The novel airborne coronavirus, termed severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2), is a causative agent of global pandemic declared by WHO in March 2020. Elucidation of protein –protein interactions (PPI) between pathogen and host is a viable paradigm in understanding protein functions and subsequently providing comprehensive knowledge for antiviral drug targeting. In this studty we used a eukaryotic CFPS system based on Leishmania tarentolae extract (LTE) in combination with AlphaLISA technology to identify intra-viral interacting proteins and putative viral-human protein interactions that can be targeted by existing or novel anti-viral therapeutics. Among 57 human-viral protein pairs, 45 binary protein interaction were identified and their direct interaction in this in vitro system was confirmed. In addition, this platform is applicable to perform antibody screening against SARS-CoV-2 RBD domain.