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. 2021 Dec 22;38(3):110242. doi: 10.1016/j.celrep.2021.110242

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© 2021 The Author(s)

Since January 2020 Elsevier has created a COVID-19 resource centre with free information in English and Mandarin on the novel coronavirus COVID-19. The COVID-19 resource centre is hosted on Elsevier Connect, the company's public news and information website. Elsevier hereby grants permission to make all its COVID-19-related research that is available on the COVID-19 resource centre - including this research content - immediately available in PubMed Central and other publicly funded repositories, such as the WHO COVID database with rights for unrestricted research re-use and analyses in any form or by any means with acknowledgement of the original source. These permissions are granted for free by Elsevier for as long as the COVID-19 resource centre remains active.

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An integrated workflow for interrogating the antibody specificity of PCs from convalescent patients with COVID-19

Serum and PBMCs are collected from patients (with confirmed SARS-CoV-2 PCR positive test). The serum is assayed with IgA and IgG ELISAs as well as POCTs. From a subset of 16 patients, PCs are isolated from PBMCs by magnetic cell sorting and to then undergo gel encapsulation and barcoding for single-cell sequencing of their antibody heavy (HC) and light chain (LC) transcripts. Antibody repertoire analysis is performed to identify expanded PC clonal lineages, which are then reformatted into single open reading frame (ORF) full-length synthetic antibody genes, including homology arms, to allow for single-step cloning-free genome editing. The resulting mammalian display library then undergoes high-throughput screening for SARS-CoV-2 binding by flow cytometry and deep sequencing to recover the identity of the corresponding clonal lineages. Supernatant is used to determine the cross-reactivity of antibodies within the library with coronavirus antigens.