Protocol to isolate broadly neutralizing monoclonal antibodies against SARS-CoV-2 from human B cells.

Protocol to isolate broadly neutralizing monoclonal antibodies against SARS-CoV-2 from human B cells.

Publication date: Dec 12, 2025

Broadly neutralizing monoclonal antibodies (bnAbs) against SARS-CoV-2 are crucial for developing therapeutics and vaccines that confer broad protection against SARS-CoV-2 variants and related sarbecoviruses. Here, we present a protocol to isolate bnAbs against SARS-CoV-2 from human B cells. We describe steps for sorting single spike-specific B cells from human peripheral blood mononuclear cells, cloning monoclonal antibodies (mAbs) from single B cells, screening mAbs with broadly neutralizing activity, and measuring the neutralizing potency of bnAbs against a panel of pseudoviruses. For complete details on the use and execution of this protocol, please refer to Chen et al. .

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Concepts Keywords
Antibodies Antibody
Complete High-throughput screening
Execution Immunology
Sarbecoviruses Microbiology
Therapeutics Molecular biology
Single cell

Semantics

Type Source Name
disease MESH STAR
disease MESH infection
drug DRUGBANK Edetic Acid
disease MESH PBS
disease MESH FBS
drug DRUGBANK Coenzyme M
drug DRUGBANK Nitrogen
drug DRUGBANK Glycerin
drug DRUGBANK Ampicillin
drug DRUGBANK Methylergometrine
disease MESH SMS
drug DRUGBANK Sodium lauryl sulfate
disease MESH SDS
drug DRUGBANK Water
drug DRUGBANK 2′-deoxyuridylic acid
disease MESH APC
drug DRUGBANK Immune Globulin Human
disease MESH strains
drug DRUGBANK Glycine
drug DRUGBANK Phosphate ion
drug DRUGBANK Trypsin
drug DRUGBANK Streptomycin
drug DRUGBANK Ethanol
drug DRUGBANK Sodium Chloride
drug DRUGBANK Tromethamine
drug DRUGBANK Fosfomycin
disease MESH FCM
disease MESH Rad
drug DRUGBANK Dextrose unspecified form
disease MESH ddH
disease MESH rupture
drug DRUGBANK Aspartame
drug DRUGBANK Medical air
pathway REACTOME Digestion
disease MESH shock
drug DRUGBANK Esomeprazole
disease MESH Plan
drug DRUGBANK Hepatitis B Vaccine (Recombinant)

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