Abstract
After the pandemic of COVID-19, neutralizing antibodies (NAbs) against SARS-CoV-2 have been developed for the prophylactic and therapeutic purposes. However, few methodologies are described in detail on how to rapidly and efficiently generate effective NAbs to SARS-CoV-2. Here, we integrated and optimized a strategically screening method for NAbs, which has enabled us to obtain SARS-CoV-2 receptor-binding domain (RBD) specific NAbs within 6 days, followed by additional 9 days for antibody production and function analysis. Using this method, we obtained 198 specific Abs against SARS-CoV-2 RBD from the blood samples of COVID-19 convalescent patients, and 96 of them showed neutralizing activity. At least 20% of these NAbs exhibited advanced neutralizing potency and high affinity, with the top two NAbs showing half-maximal inhibitory concentration (IC50) to block authentic SARS-CoV-2 at 9.88 and 11.13 ng/ml, respectively. Altogether, our study provides an effective methodology with high applicable value for discovering potential preventative and therapeutic NAbs for the emerging infectious diseases.
Highlights
In the past two decades, pandemic outbreaks of three novel pathogenic human coronaviruses, severe acute respiratory syndrome coronavirus (SARS-CoV), Middle East respiratory syndrome coronavirus (MERS-CoV) and SARS-CoV-2, have caused high mortality and unprecedented social and economic consequences [1,2,3,4]
Linear expression cassettes were assembled by overlapping PCR [25], which contained the essential elements for Ab gene transcription, including the CMV promoter, the antibody variable region, the antibody constant region, and the poly(A)-tail (Supplementary Table 2)
Supernatants of the transfected cells were evaluated for the S and receptor-binding domain (RBD) specific binding activities by enzyme-linked immunosorbent assay (ELISA) at Day 4, and their neutralizing capacities to pseudovirus were tested at Day 6
Summary
In the past two decades, pandemic outbreaks of three novel pathogenic human coronaviruses, severe acute respiratory syndrome coronavirus (SARS-CoV), Middle East respiratory syndrome coronavirus (MERS-CoV) and SARS-CoV-2, have caused high mortality and unprecedented social and economic consequences [1,2,3,4]. SARS-CoV-2 Neutralizing Antibodies diseases, NAbs are an alternative treatment strategy for blocking virus infection. During the outbreaks of SARS-CoV and MERSCoV, convalescent patients’ plasma containing NAbs was a safe and effective treatment option to reduce mortality in severe cases [5, 6]. Convalescent plasma is limited and the polyclonal non-neutralizing Abs in the plasma may cause undesired side effects [7]. The NAb therapeutics is an effective replacement of convalescent plasma therapy. A rapid and efficient screening method of desired NAbs is greatly needed
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