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