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Summary: Prior to receptor engagement, a specific, non-electrostatic glycan-glycan interaction between viral particles and host cells may lengthen the dwell time of the virus at the cellular surface, thereby facilitating subsequent virus entry. Here, we present a protocol for quantifying the level of glycan-mediated binding between virus or virus-like-particles and human peripheral blood mononuclear cells (PBMCs) using a nanoluciferase reporter system. We describe steps for virus production, isolation of PBMCs, and performing a nanoluciferase binding assay.For complete details on the use and execution of this protocol, please refer to Spillings et al.1 : Publisher’s note: Undertaking any experimental protocol requires adherence to local institutional guidelines for laboratory safety and ethics.