SARS-CoV-2 Neutralization Assay Protocol

TZM-bl neutralization assays were performed as previously described (41 (link), 43 (link), 44 (link)). Briefly, antibody titrations were incubated with pseudotyped viruses for 1 h at 37°C. TZM-bl cells were diluted in DMEM to 100,000 cells/ml and added to the virus/inhibitor mix. Cells were then incubated for 48 h at 37°C. Viral entry was determined by luciferase readout with BriteLite Plus (Perkin Elmer, Waltham, MA) and read on a Victor X3 plate reader (Perkin Elmer, Waltham, MA).

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Davis-Gardner M.E., Alfant B., Weber J.A., Gardner M.R, & Farzan M. (2020). A Bispecific Antibody That Simultaneously Recognizes the V2- and V3-Glycan Epitopes of the HIV-1 Envelope Glycoprotein Is Broader and More Potent than Its Parental Antibodies. mBio, 11(1), e03080-19.

Publication 2020

BriteLite Plus Victor X3 plate reader

Antibody Assays Cells Luciferase Pseudotyped viruses Titrations Viral entry Virus

Corresponding Organization :

Other organizations : Scripps Research Institute

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Variable analysis

independent variables

Antibody titrations

dependent variables

Viral entry

control variables

Incubation time (1 h at 37°C)
Cell line (TZM-bl cells)
Cell density (100,000 cells/ml)
Incubation time (48 h at 37°C)
Luciferase readout using BriteLite Plus and Victor X3 plate reader

controls

Positive control: Not explicitly mentioned
Negative control: Not explicitly mentioned

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