Efficient generation of human anti-cytomegalovirus IgG monoclonal antibodies from preselected antigen-specific B cells.
Human anti-cytomegalovirus (CMV) specific B cells were enriched to a purity of up to 38% on CMV-coated dishes and subsequently clonally expanded in the presence of human T cell supernatant and irradiated murine thymoma helper cells. The expanded cells were immortalized by a mini-electrofusion with K6H6B5 myeloma cells. Twenty-two anti-CMV positive B cell clones could be obtained from as little as 1.5 ml donor blood. The majority of these clones produced anti-CMV antibodies of the IgG class. Ten anti-CMV positive B cell clones were submitted to separate mini-electrofusions yielding stable, human anti-CMV IgG-producing hybridomas in six out of ten fusions. These antibodies recognized different proteins of the CMV virus, as deduced from the immunofluorescence staining pattern on infected human fibroblasts.
Steenbakkers
P G
PG
Department of Immunology, Organon International b.v., Oss, The Netherlands.
Van Wezenbeek
P M
PM
van Zanten
J
J
The
T H
TH
D000911:Antibodies, Monoclonal / Q000096:biosynthesis*; D000914:Antibodies, Viral / Q000096:biosynthesis*; D001402:B-Lymphocytes / Q000276:immunology*; D003587:Cytomegalovirus / Q000276:immunology*; D000939:Epitopes; D006801:Humans; D007074:Immunoglobulin G / Q000096:biosynthesis*; D008213:Lymphocyte Activation
D000911:Antibodies, Monoclonal; D000914:Antibodies, Viral; D000939:Epitopes; D007074:Immunoglobulin G