Author: Xin Zeng; Lingfang Li; Jing Lin; Xinlei Li; Bin Liu; Yang Kong; Shunze Zeng; Jianhua Du; Huahong Xiao; Tao Zhang; Shelin Zhang; Jianghai Liu
Title: Blocking antibodies against SARS-CoV-2 RBD isolated from a phage display antibody library using a competitive biopanning strategy Document date: 2020_4_20
ID: 4w6caxwu_15
Snippet: The copyright holder for this preprint (which was not peer-reviewed) is the . https://doi.org/10.1101/2020.04.19.049643 doi: bioRxiv preprint Phage ELISA Single clones were inoculated into 400μl 2×YT medium containing 50μg/ml carbenicillin, 25μg/ml kanamycin and 10 10 pfu/ml helper phages in 96-deep-well plates and incubated overnight at 37°C and 250rpm. The plates were centrifuged at 4,000rpm and the supernatant was applied for phage ELISA......
Document: The copyright holder for this preprint (which was not peer-reviewed) is the . https://doi.org/10.1101/2020.04.19.049643 doi: bioRxiv preprint Phage ELISA Single clones were inoculated into 400μl 2×YT medium containing 50μg/ml carbenicillin, 25μg/ml kanamycin and 10 10 pfu/ml helper phages in 96-deep-well plates and incubated overnight at 37°C and 250rpm. The plates were centrifuged at 4,000rpm and the supernatant was applied for phage ELISA. The 96-well Maxisorp plates were coated overnight at 4°C with RBD-mFc (1μg/ml, 100μl per well). After blocking with 1% PVA, plates were incubated with 50μl bacterial supernatant containing phages for 2 hours at room temperature. After six times of wash with PT, bound phages were detected using an HRP-conjugated anti-M13 antibody (Sino biological) and tetramethyl benzidine (TMB) as substrate. Absorption at 450nm was measured.
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