Abstract
Passive serotherapy can confer immediate protection against microbial infection, but methods to rapidly generate human neutralizing monoclonal antibodies are not yet available. We have developed an improved method for Epstein-Barr virus transformation of human B cells. We used this method to analyze the memory repertoire of a patient who recovered from severe acute respiratory syndrome coronavirus (SARS-CoV) infection and to isolate monoclonal antibodies specific for different viral proteins, including 35 antibodies with in vitro neutralizing activity ranging from 10(-8)M to 10(-11)M. One such antibody confers protection in vivo in a mouse model of SARS-CoV infection. These results show that it is possible to interrogate the memory repertoire of immune donors to rapidly and efficiently isolate neutralizing antibodies that have been selected in the course of natural infection.
Publication types
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Research Support, Non-U.S. Gov't
MeSH terms
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Adult
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Animals
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Antibodies, Monoclonal / isolation & purification*
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B-Lymphocytes / immunology*
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Chlorocebus aethiops
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DNA Primers
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Disease Models, Animal
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Enzyme-Linked Immunosorbent Assay
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Herpesvirus 4, Human / metabolism
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Humans
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Immunization, Passive / methods*
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Immunologic Memory / immunology*
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Membrane Glycoproteins / genetics
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Membrane Glycoproteins / metabolism
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Mice
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Microscopy, Immunoelectron
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Neutralization Tests
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Polymerase Chain Reaction
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Severe Acute Respiratory Syndrome / immunology*
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Spike Glycoprotein, Coronavirus
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Transformation, Genetic
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Vero Cells
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Viral Envelope Proteins / genetics
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Viral Envelope Proteins / metabolism
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alpha-Macroglobulins / metabolism
Substances
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Antibodies, Monoclonal
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DNA Primers
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Membrane Glycoproteins
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Spike Glycoprotein, Coronavirus
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Viral Envelope Proteins
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alpha-Macroglobulins
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polyclonal B cell activator
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spike glycoprotein, SARS-CoV