Establishment of a well-characterized SARS-CoV-2 lentiviral pseudovirus neutralization assay using 293T cells with stable expression of ACE2 and TMPRSS2

Pseudoviruses are useful surrogates for highly pathogenic viruses because of their safety, genetic stability, and scalability screening assays. Many different pseudovirus platforms exist, each with advantages limitations. Here we report our efforts to optimize characterize an HIV-based lentiviral assay neutralizing antibodies SARS-CoV-2 using a stable 293T cell line expressing human angiotensin converting enzyme 2 (ACE2) transmembrane serine protease (TMPRSS2). We assessed target cells, established conditions that generate readouts over at least two-log range, confirmed consistent neutralization titers range input. Using reference sera plasma panels, evaluated precision showed correlate well results reported in other Overall, is relatively simple, scalable, suitable variety entry